Avian immunoglobulin Y immunological response to the major peanut allergens

Avian immunoglobulin Y immunological response to the major peanut allergens | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Avian immunoglobulin Y immunological response to the major peanut allergens Kanaka Durga Devi Nelluri, Anupama Ammulu Manne, Praveen Kumar Vemuri, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7161972/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Introduction Specifically in animal models of human infectious illnesses, chicken immunoglobulin Y (IgY), a highly conserved counterpart of human immunoglobulin G (IgG), has demonstrated advantages and a favourable safety profile. IgY is inexpensive, quick-acting, and simple to make. Egg-laying chickens can easily produce huge quantities of IgY antibodies without damaging the environment or infrastructural expenditure. Over 30% of people suffer from immunoglobulin E (IgE)-mediated allergy, which is the most prevalent hypersensitivity illness. Althoughabout 1%-2% of the general population suffers from peanut allergy, peanuts are one of the most pervasive food causes of deadly anaphylaxis globally. Results Arachis hypogea contains allergens that exhibit immunological responses towards IgY antibodies. Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS PAGE) analysis was performed to check the purity of extracted peanut and IgY samples, acquiring the molecular weight patterns of heavy and light chains of 75 and 18 kDa. The peanut proteins were screened for their activity against immunoglobulins for allergic responses through the Enzyme-linked Immunosorbent assay (ELISA) technique. Among the white hen, brown hen and duck eggs, the white eggs' concentration of IgY antibodies showed an excellent sensitivity with increased attention to the Arachis hypogea antigens.FTIR analysis of peanut proteins was carried out to identify extracted proteins. Antigen-antibody interaction was confirmed through immunoblotting. We observed distinct banding patterns for the peanut allergens Ara h1, Ara h3, and Ara h6 when probed with specific monoclonal antibodies. Conclusion Designing therapeutic IgY antibodies that can passively suppress IgE-mediated reactions in allergy patients is another potential future approach. Arachis hypogea ELISA FTIR IgY SDS PAGE Western blotting Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction The peanut ( Arachis hypogaea L.) along with its wild cousins are native to South America, which are previously been found in Bolivia, Argentina, Paraguay, Uruguay, and present-day Brazil. An uncommon hybridization occurrence between two diploid species, Arachis duranensis (AA genome) and Arachis ipaensis (BB genome), produced the tetraploid (AABB genome) cultured peanuts that are grown today. They belong to the legume family (Fabaceae) 1 . Many potent allergenic proteins in peanuts can cause serious anaphylaxis reactions, even in small doses. A sizable segment of the US populace is in contact with peanuts. However, it was shown that 2.1% of children under the age of 18 and 1.4% of adults in the US generally suffer from peanut and tree nut allergies. While peanut allergy sufferers experience peanut-related anaphylaxis very frequently, fatalities are still extremely uncommon. Allergen immunotherapy, which has been used for over a century and is usually given as subcutaneous injections of soluble allergen extracts, is an effective treatment for many IgE-mediated allergies 2 . Subcutaneous injections containing peanut extract were explored more than 20 years ago. Despite their effectiveness in treating allergies to insects and the environment, the technique was abandoned because of the high rate of anaphylaxis that followed injections 3 . In the last ten years, researchers have changed the way allergen immunotherapy is administered to treat food allergies to reduce side effects while continuously exposing allergic patients to the allergen that causes their symptoms. These antigen delivery methods include ingesting the allergen orally (oral immunotherapy; OIT), administering a soluble protein extract sublingually (sublingual immunotherapy; SLIT), and applying dry allergens topically (epicutaneous immunotherapy; EPIT). Blocking Fel d1's binding spots, or epitopes prevents it from binding with IgE and, thus, from inducing an allergic reaction, which is another method of easing allergy symptoms. This is just one of the ways allergen-specific immunotherapy (ASIT) works to increase the afflicted person's anti-Fel d1 IgG antibodies production. These particular IgG antibodies attach to Fel d1, blocking its IgE binding and, consequently, the allergic response 4 . Although ASIT has been utilized in allergy treatment and has a complex mechanism of action, ASIT has several disadvantages, including cost, reaction time, and potential side effects. It has been shown that anti-Fel d1-specific polyclonal immunoglobulin Y (sIgY) in chicken egg yolks counteracts the allergic effects of Fel d1 mast cell culture techniques 4 . The avian immunoglobulin known as IgY is comparable to mammalian IgG. To give passive immunity to their young ones, hens naturally create these antibodies in response to antigen exposure. These antibodies are then transported and concentrated into egg yolks. Therefore, large amounts build up in the yolks of chicken eggs, which can be removed, refined, and used to make food. By attaching to active binding sites on the targeted proteins, these antibodies successfully lessen the antigenicity and allergenicity of those proteins. Oral administration of antibodies derived from chicken eggs has been shown in numerous studies to be both safe and effective in reducing diarrhoea in domestic animals 5 . Eggs naturally contain IgY antibodies, which are generally not harmful. They also produce more quickly and at a lower cost than vaccinations and may be kept in powder form at low moisture for months without refrigeration. There are other general benefits to using IgY antibodies in immunotherapy. From an animal welfare view, egg collection is non-painful and kind to the animals. Furthermore, compared to mammalian IgG, IgY has a higher binding avidity to target antigens. Because of the evolutionary difference between mammals and birds, IgY antibodies can be generated against conserved mammalian proteins more readily and effectively than IgG antibodies in other animals. Moreover, fewer antigens are required for an effective immune response 6 . Protein molecules called antibodies are created in response to an antigen. They are frequently employed in research, diagnosis, and therapy because of their capacity to bind to specific targets. Most of the antibodies on the market are made by animals, particularly tiny rodents 7 . However, particular antigens elicit modest immune responses or are even entirely non-immunogenic, making generating antibodies in mammals difficult. Additionally, the process through which mammals produce antibodies necessitates the inoculation, blood sample collection, and slaughter of animals 8 . Egg yolk antibodies (IgY) are gaining popularity as researchers look for methods that are more effective, affordable, and refined in their usage of animals. Additionally, the percentage of particular IgY generated by hens ranges from 1 to 10% of all antibodies 9 . However, the quantity of antigen administered to the hen and its immunogenicity and molecular weight are connected to the amount of antibodies generated 10 . IgY technology has been employed for numerous human and veterinary health objectives due to its productive nature and more refined approach to animal welfare 11 . The Arachis hypogea , or peanut plant, is a bean/legume family Fabaceae member 12 . Even small amounts of peanut allergens can trigger symptoms, ranging from mild reactions such as itching to more severe manifestations like anaphylaxis, which is life-threatening. The peanut plant, scientifically known as A. hypogea , contains several allergens that can stimulate the specific IgE antibodies production in individuals prone to peanut allergens 13 . Distinct populations in diverse geographic regions can have different sensitivity patterns to these allergies 14 . The major allergens are Ara h 1, Ara h 2, Ara h 3 and Ara h 6. Particularly severe allergy responses are linked to some allergens 15 . Peanuts have two bicupin seed storage proteins, Ara h 1 and Ara h 3. They belong to the highly functionally varied protein group known as the cupin superfamily 16 . Peanuts also contain other seed storage proteins known as Ara h 2, Ara h 6 and Ara h 7. These proteins are seed storage proteins as of the prolamin superfamily's 2S albumin class 17 . Prolamins are a collection of plant storage proteins commonly found in cereal grains such as wheat, barley and maize. In this research, the yolks of white, brown hen, and duck eggs, IgY antibodies were isolated. SDS PAGE helps to check the purity of extracted proteins and IgY antibodies. The immune activity of peanut antigen towards IgY antibodies from egg yolks was demonstrated through ELISA. Western blotting was performed to confirm antigen-antibody reaction. Materials and Methods Extraction and Purification of IgY from chicken and duck eggs Different varieties of chicken eggs, such as white and brown eggs and duck eggs, are taken, and the yolk is parted from the white. The yolk is then put into a 50 ml tube, and its volume is recorded 18 . Phosphate buffer saline (PBS) is added twice the egg yolk volume, and then 3.5% polyethylene glycol (PEG 6000) of the entire volume is added and vortexed for ten minutes. The samples undergo a 20-minute, 10,000 rpm centrifugation at 4 0 C. After filtering the supernatant and transferring it to a new tube, 8.5% PEG 6000 (estimated based on the revised volume) is added and vortexed as previously described. Once more, the samples are centrifuged at 4 0 C for 20 minutes at 10,000 rpm. The pellet is then meticulously dissolved in 1 ml of PBS with the help of a glass stick or vortexer, and the volume of PBS is increased to 10 ml. The sample is combined with 12% (w/v) PEG 6000 and vortexed similarly. Using a glass stick or vortexer, the pellet is thoroughly dissolved in 800 µl of PBS after the samples are centrifuged for 20 minutes at 10,000 rpm at 4 0 C. After transferring the sample into a dialysis tube, rinsed it with 400 µl PBS and introduced it to the dialysis machine. The extract is dialyzed in 0.1% saline using a magnetic stirrer and slowly swirled all night. PBS was used in place of the saline the next day, and dialyzing was done for an additional three hours. The finished samples are examined on SDS-PAGE and kept at -20 0 C 19 . SDS PAGE confirmation of IgY Using SDS-PAGE gel electrophoresis, the isolated IgY’s molecular weight was assessed 20 . It was completed in a vertical gel slab system. Each well received 10 mL of each sample, which was then migrated for two hours at a constant 100 V. After migration, gels were dyed using Coomassie Brilliant Blue R-250. Soluble peanut protein extraction from peanut flour Lightly roasted and finely ground peanut flour was prepared. PBS was combined with peanut flour in a 1:4 (w/v) ratio 21 . The mixture was undergone centrifugation at 15,000 rpm for 45 minutes at 4 0 C after being agitated for 1.5 h with 6 M NaOH to maintain a constant pH of 8.5. The supernatant of the protein extraction was filtered through dialysis. The dialysis membrane was soaked in water for 10–15 minutes before incorporating the sample. The filled Dialysis membrane was immersed in 10 mM phosphate buffer, and using the magnetic stirrer the sample was stirred for 2 h. The extract was diluted with PBS for SDS-PAGE analysis. SDS-PAGE analysis of purified peanut proteins A volume of 10 ul was added to the loading buffer and incubated at 100 0 C for 8 minutes. When the gel loading dye reached the end of the gel, electrophoresis was conducted for 2 h at 100V with samples loaded along with a medium-range prepared protein marker. Gels remained dyed with Coomassie Brilliant Blue R-250 following migration. Destaining was done in 25% methanol, 10% acetic acid and 65% distilled water 22 . FTIR analysis of proteins A Bruker Tensor 27 FTIR spectrometer (Bruker, Germany) was used to collect the spectra 23 . In a wavenumber range of 4000 cm − 1 to 400 cm − 1 , spectra of each sample were collected at room temperature with a resolution of 4 cm − 1 . Normalization of data is done by the software named OPUS. Matlab 2014b (Mathworks, Natick MA, USA) was used by in-house software to carry out the final analysis 24 . Immunological activity of IgY towards peanut antigens To determine the allergen's selectivity towards IgY, Affymetrix eBioscience 96-well plates coated with a 10 µg/ml allergen concentration and incubated in the room for two hours 25 . 200 µl of blocking buffer was added to each well after three rounds of washing with 1X Tris-buffered saline and 0.1% Tween® 20 detergent (TBST) wash buffer. The wells were then left to remain at room temperature for an hour. The primary antibodies (IgY) were loaded and incubated for 30 minutes at room temperature. The mixing of a solution containing a secondary antibody (HRP conjugated Goat anti-chicken IgY antibody) (Santa-Cruz)) Conjugation to an enzyme detects the attached antibody after unbound antibodies are washed away. Unbound conjugate is removed after incubation, and 100 µl of the substrate solution (TMB, eBiosciences) is added. Utilizing an ELISA plate reader, substrate converted to coloured product is measured. Western blotting SDS-PAGE was performed using BioRad SDS-PAGE equipment in line with the directions in the GeNei Western Blotting Teaching Kit. The resultant gel was rinsed three times with distilled water at room temperature before being fixed for 15 minutes in a solution of acetic acid, isopropanol, and water (10:25:65). According to the manufacturer's suggestions, the gel was sandwiched inside the GeNei blotting cassette to transfer bands onto the nylon membrane and electrophoresed at 50V for two hours 26 . The membrane was carefully removed from the cassette and submerged in 10 ml of newly made blocking buffer for the night at 4 0 C. The following day, the buffer was thrown away, and the membrane was washed thrice with 1X PBST (phosphate-buffered saline, 0.05% Tween-20) while being incubated for 3 minutes. After that, the membrane was incubated for 30 minutes at room temperature with light shaking in 10 ml of primary IgY antibody solution. After a second round of cleaning as directed, the membrane was incubated with 10 ml of horseradish peroxidase-conjugated anti-chicken IgG for 30 minutes at room temperature with light shaking. The alkaline phosphates substrate solution in 10 ml was incubated while being gently stirred for five to ten minutes to check for the intensity of the coloured band after the wash procedure was completed four times. The membrane was tested after being washed with distilled water, and the findings were noted. Results SDS-PAGE confirmation of IgY For the pure IgY of white hen eggs, SDS-PAGE analysis showed two major bands with heavy and light chains. The heavy chain with 75 kDa and the light chain with 18 kDa was confirmed through SDS-PAGE (Fig. 1 ). The rest of the two eggs (brown hen egg and duck egg) contain more IgY-like proteins, which may act as contaminants or inhibitors during antigen-antibody interactions. As white hen eggs showed the presence of pure IgY, it was chosen for further study. SDS-PAGE analysis of peanut proteins All protein concentrations of 1 mg/ml in SDS sample buffer were adjusted before same amounts of total protein (8 µg) were added into each sample for the analysis. SDS-PAGE was conduceted to confirm the presence of peanut antigens in the extracted samples (Fig. 2 ). Ara h 1, Ara h 2, Ara h 3 acidic form, Ara h 3 basic form, Ara h 6 are present in peanut extracts. Ara h 3 basic structure is more prominent in after dialysis sample. FTIR analysis of peanut proteins FTIR spectroscopy of peanut antigens before and after dialysis can reveal details about the modifications in molecule structure and composition throughout the dialysis procedure. Before dialysis, the FTIR spectrum of peanut antigens exhibited distinctive absorption peaks corresponding to the numerous functional groups, including amide bonds, carbohydrates, lipids, and nucleic acids, present in the antigens. The spectrum may also point to some peanut allergens that can cause allergy reactions in sensitive people, including Ara h 1, Ara h 2, Ara h 3, and Ara h 6 (Fig. 3 ). ELISA for IgY against antigen ELISA experiment was performed using an increasing concentration of IgY antibodies with a constant concentration of peanut antigen (Fig. 4 ). The maximum absorbance was identified at 500 mg/ml of IgY antibody for all three eggs. Western blotting The Genetix transblot device was used to blot proteins onto 0.2 µm nitrocellulose membranes for two hours at 125 mA (Fig. 5 ). Then the membranes were probed with a specific monoclonal antibody to epitopes on purified IgY (primary antibody) overnight at 4 0 C. An enzyme-conjugated secondary antibody was coupled to the membranes for an hour following washing. Immunoblot analysis of a peanut sample probed with specific antibodies against major peanut allergens, including Ara h1, Ara h3, and Ara h6. Primary antibodies against Ara h1, Ara h3, and Ara h6 were added to the peanut extract after it had been separated by gel electrophoresis and moved on a membrane. Discussion IgY technology is a rapidly expanding subject in the biological sciences nowadays. IgY's in body lotions, cosmetics, yoghurt, and other functional foods, as well as powder-form supplements, may be able to prevent or treat human diseases. Reducing the dosage of antibiotics used to treat bacterial infections of the digestive tract is possible with IgY antibodies. Furthermore, egg yolk antibodies offer a fresh strategy for treating intestinal parasites and Candida albicans 27 . SDS-PAGE was used to evaluate the fractions obtained after IgY extraction (Fig. 1 ). The projected molecular masses were matched by the molecular weight patterns for the heavy and light chains, which were 75 and 18 kDa, respectively 28 . The strength of the Ara h 1 (62 kD) monomer band was noted based on the protein ladder (Fig. 2 ). In the peanut protein sample-buffer-soluble fractions of the SDS, the aggregated protein bands were more noticeable (Fig. 2 ). The outcome was consistent with a few previous research investigations 29 . As the peanuts were roasted, strong connections were found between aggregate formation seen in Fig. 2 and decreased in water-soluble Ara h1. This suggested that heat processes may lead to the water-soluble proteins being broken down by free radicals attacking the peptide bonds and side chains of the proteins, or they could induce the proteins to assemble by chemical cross-linking. Birds have a strong immunological response to mammalian antigens because of the evolutionary separation between mammals and birds. This improves IgY's binding specificity, as demonstrated by immunological diagnostic techniques such as immunohistochemistry, ELISA, and immunofluorescence 27 . This may be attributed to a loss of IgY binding because of structural changes in peanut proteins, as opposed to brown and duck eggs, where there was a low detection of IgY concentration following an increase in protein concentration (Fig. 4 ). In white eggs, the concentration of IgY antibodies showed a good sensitivity with an increase in the concentration of the Arachis hypogea antigens. A fascinating investigation was conducted on the pro-carcinogenic Helicobacter pylori infection, which affects the stomach of humans and emits vacuolating cytotoxin A (VacA), which promotes the bacteria's persistence. There have been reports that oral administration of IgY against this toxin minimizes the histological alterations generated in the stomach tissue and protects against Helicobacter pylori infection 30 . Since the secondary structure of proteins is associated with the amide I band (1600–1700 cm − 1). It was chosen for further study because it is the most sensitive vibrational band of the protein backbone. According to previous studies, the bands that corresponded to the secondary structures 31 . The α-helix was represented by the bonds at 1650–1660 cm − 1, the β-sheet by 1618–1640 and 1670–1690 cm − 1, the β-turn by 1660–1670 and 1690–1700 cm − 1, and the random coils by 1645 cm − 1. The band areas of linked peaks were used to calculate the percentages of every single secondary structural component. The dialysis process aims to remove small molecules and impurities, so a reduction in the intensity of peaks associated with these impurities might be observed. However, the main structural features of the peanut antigens, including the characteristic absorption peaks, are not expected to be significantly altered by dialysis alone. Peanut protein extract transmitted to a membrane, and probed with a specific monoclonal antibody against Ara h1, Ara h3 and Ara h6. Following primary and secondary antibody incubations, the blot was developed using chemiluminescence, and the resulting bands were analyzed for the presence and molecular weight of allergic proteins. This approach provides insights into the Ara h1’s specificity and expression in the peanut protein extract. We observed distinct banding patterns for the peanut allergens Ara h1, Ara h3, and Ara h6 when probed with specific monoclonal antibodies. The monoclonal antibody against Ara h1 revealed a prominent band at 64 KDa, consistent with the expected size of Ara h1. When probing for Ara h3, a distinct band at 48 KDa was detected, indicating the presence of Ara h3 in the peanut protein extract. Similarly, the monoclonal antibody against Ara h6 identified a specific band at 14.5 KDa, confirming the presence of Ara h6. These results suggest the co-existence of Ara h1, Ara h3, and Ara h6 in the peanut sample, providing valuable insights into the allergenic protein composition. In an in vivo skin infection model, the IgY immunophototherapy was very specific, efficacious, and did not harm the healthy epithelium. By changing only the specific antibody, this innovative method of eliminating germs specifically could be utilized for anti-infection treatment, perhaps decreasing the adversative effects of antibiotic use due to selection for resistant variations 32 . Conclusions Peanut proteins can be investigated as a significant source of products that have immunological responses. The ELISA test examined the antigenicity. The present work established that among the three eggs chosen, the concentration of IgY antibodies in the white egg shows a good sensitivity with an increase in the concentration of the peanut antigens. Western blotting was performed to confirm that this is the same protein acting as antigen or allergen during an immunological response. However, SDS-PAGE analysis of peanut proteins confirmed the presence of peanut antigens in the extracted sample. Western blotting confirmed the activity of Ara h3 towards IgY antibody. The IgY's successful performance encourages further research into its in vivo efficacy and clinical trials against A. hypogea . Thanks to their tremendous potential, igY antibodies have a bright future in the treatment of many different diseases. Abbreviations IgG immunoglobulin G IgY immunoglobulin Y IgE immunoglobulin E SDS PAGE Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis ELISA Enzyme-linked Immunosorbent assay PBS Phosphate buffer saline PEG 6000 polyethylene glycol TBST 1X Tris-buffered saline with 0.1% Tween® 20 detergent 1X PBST phosphate-buffered saline 0.05% Tween-20. Declarations -Ethics approval and consent to participate I/We Kanaka Durga Devi.N, Anupama .A. Manne, Praveen Kumar Vemuri, Tanvitha Vadlamudi, Prathyusha Chowdary Koduri, Vaka koti reddy, Kallm Dhanvanth reddy, the author(s) of manuscript entitled “ Avian immunoglobulin Y immunological response to the major peanut allergens ” do hereby authorize you to publish the above said manuscript in the journal. I/We further state that: This article/Paper authored by me/us is an original and genuine work. It does not infringe on the right of others and does not contain any libelous or unlawful statement. It has neither been submitted for review/publication nor published elsewhere in any print/electronic form. The tables, figures or data are not copied from anyother publications. I/We permit authorities concerned to publish the said paper in the journal with editorial modifications, if any. -Consent for publication I/We Kanaka Durga Devi.N, Anupama .A. Manne, Praveen Kumar Vemuri, Tanvitha Vadlamudi, Prathyusha Chowdary Koduri, Vaka koti reddy, Kallm Dhanvanth reddy, the author(s) of manuscript entitled “ Avian immunoglobulin Y immunological response to the major peanut allergens ” do hereby authorize you to publish the above said manuscript in the journal -Availability of data and material Not applicable -Competing interests The authors do not have any conflict of interest”. -Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. -Authors' contributions All authors contributed to the study conception and design. Material preparation, project administration, supervision, first draft of the manuscript was written by Manne Anupama Ammulu . Data collection, analysis, methodology, manuscript review and editing by Kanaka durga devi Nelluri. Visualization and analysis by Praveen Kumar Vemuri . Data collection and methodology, drafting manuscript by Tanvitha Vadlamudi, Prathyusha Chowdary Koduri, Vaka koti reddy, Kallm Dhanvanth reddy . All authors read and approved the final manuscript. -Acknowledgements Authors are thankful to the Principal and management of KVSR Siddhartha college of pharmaceutical sciences, Vijayawada for providing all the facilities for this research project. References Zhuang W, Chen H, Yang M. The genome of cultivated peanut provides insight into legume karyotypes, polyploid evolution and crop domestication. Nat Genet . 2019;51(5):865–876. doi: 10.1038/s41588-019-0402-2 Viswanathan RK, Busse WW. Allergen immunotherapy in allergic respiratory diseases: From mechanisms to meta-analyses. Chest . 2012;141(5):1303–1314. doi: 10.1378/chest.11-2800 Johnson-Weaver BT, Staats HF, Burks AW, Kulis MD. Adjuvanted immunotherapy approaches for peanut allergy. Front Immunol . 2018;9(SEP):1–8. doi: 10.3389/fimmu.2018.02156 Satyaraj E, Sun P, Sherrill S. Fel d1 Blocking Antibodies: A Novel Method to Reduce IgE-Mediated Allergy to Cats. J Immunol Res . 2021;2021. doi: 10.1155/2021/5545173 Satyaraj E, Li Q, Sun P, Sherrill S. Anti-Fel d1 immunoglobulin Y antibody-containing egg ingredient lowers allergen levels in cat saliva. J Feline Med Surg . 2019;21(10):875–881. doi: 10.1177/1098612X19861218 Abbas AT, El-Kafrawy SA, Sohrab SS, Azhar EIA. IgY antibodies for the immunoprophylaxis and therapy of respiratory infections. Hum Vaccines Immunother . 2019;15(1):264–275. doi: 10.1080/21645515.2018.1514224 A. Michael1,, S. Meenatchisundaram2* GP, Subbraj1 T, Selvakumaran3 R, Ramalingam4 and S. Chicken egg yolk antibodies (IgY) as an alternative to mammalian antibodies. Indian J Sci Technol . 2010;6(1):7. http://journals.sagepub.com/doi/10.1177/1120700020921110%0Ahttps://doi.org/10.1016/j.reuma.2018.06.001%0Ahttps://doi.org/10.1016/j.arth.2018.03.044%0Ahttps://reader.elsevier.com/reader/sd/pii/S1063458420300078?token=C039B 8B13922A2079230DC9AF11A333E295FCD8 Pauly D, Chacana PA, Calzado EG, Brembs B, Schade R. Igy technology: Extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation. J Vis Exp . 2011;(51):1–6. doi: 10.3791/3084 Larsson A, Campbell A, Eriksson M. Chicken antibodies are highly suitable for particle enhanced turbidimetric assays. Front Immunol . 2022;13(October):1–8. doi: 10.3389/fimmu.2022.1016781 Agurto-Arteaga A, Poma-Acevedo A, Rios-Matos D. Preclinical Assessment of IgY Antibodies Against Recombinant SARS-CoV-2 RBD Protein for Prophylaxis and Post-Infection Treatment of COVID-19. Front Immunol . 2022;13(May):1–12. doi: 10.3389/fimmu.2022.881604 Pereira EPV, van Tilburg MF, Florean EOPT, Guedes MIF. Egg yolk antibodies (IgY)and their applications in human and veterinary health: A review. Int Immunopharmacol . 2019;73(May):293–303. doi: 10.1016/j.intimp.2019.05.015 Iqbal A, Shah F, Hamayun M. Allergens of Arachis hypogaeaand the effect of processing on their detection by ELISA. Food Nutr Res . 2016;60(February). doi: 10.3402/fnr.v60.28945 Palladino C, Breiteneder H. Peanut allergens. Mol Immunol . 2018;100(March):58–70. doi: 10.1016/j.molimm.2018.04.005 Vereda A, Van Hage M, Ahlstedt S. Peanut allergy: Clinical and immunologic differences among patients from 3 different geographic regions. J Allergy Clin Immunol . 2011;127(3):603–607. doi: 10.1016/j.jaci.2010.09.010 Storni F, Zeltins A, Balke I. Vaccine against peanut allergy based on engineered virus-like particles displaying single major peanut allergens. J Allergy Clin Immunol . 2020;145(4):1240–1253.e3. doi: 10.1016/j.jaci.2019.12.007 Dunwell JM, Purvis A, Khuri S. Cupins: The most functionally diverse protein superfamily? Phytochemistry . 2004;65(1):7–17. doi: 10.1016/j.phytochem.2003.08.016 Mylne JS, Hara-Nishimura I, Rosengren KJ. Seed storage albumins: Biosynthesis, trafficking and structures. Funct Plant Biol . 2014;41(7):671–677. doi: 10.1071/FP14035 Pauly D, Chacana PA, Calzado EG, Brembs B, Schade R. Igy technology: Extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation. J Vis Exp . 2011;i(51):2–7. doi: 10.3791/3084 AKITA EM, NAKAI S. Immunoglobulins from Egg Yolk: Isolation and Purification. J Food Sci . 1992;57(3):629–634. doi: 10.1111/j.1365-2621.1992.tb08058.x Kirley TL, Norman AB. Unfolding of IgG domains detected by non-reducing SDS-PAGE. Biochem Biophys Res Commun . 2018;503(2):944–949. doi: 10.1016/j.bbrc.2018.06.100 Berglund JP, Szczepanski N, Penumarti A. Preparation and Analysis of Peanut Flour Used in Oral Immunotherapy Clinical Trials. J Allergy Clin Immunol Pract . 2017;5(4):1098–1104. doi: 10.1016/j.jaip.2016.11.034 Nowakowski AB, Wobig WJ, Petering DH. Native SDS-PAGE: High resolution electrophoretic separation of proteins with retention of native properties including bound metal ions. Metallomics . 2014;6(5):1068–1078. doi: 10.1039/c4mt00033a Miller LM, Bourassa MW, Smith RJ. FTIR spectroscopic imaging of protein aggregation in living cells. Biochim Biophys Acta - Biomembr . 2013;1828(10):2339–2346. doi: 10.1016/j.bbamem.2013.01.014 Strych U, Willson RC. Purification and Characterization of Proteins. Man Ind Microbiol Biotechnol . 2014;10(4):731–742. doi: 10.1128/9781555816827.ch52 Hnasko RM, Lin A V., McGarvey JA, Mattison CP. Sensitive and selective detection of peanut allergen Ara h 1 by ELISA and lateral flow immunoassay. Food Chem . 2022;396(July):133657. doi: 10.1016/j.foodchem.2022.133657 Mishra M, Tiwari S, Gomes A V. Protein purification and analysis: Next generation western blotting techniques. Expert Rev Proteomics . 2017;14(11):1037–1053. doi: 10.1080/14789450.2017.1388167 Müller S, Schubert A, Zajac J, Dyck T, Oelkrug C. IgY antibodies in human nutrition for disease prevention. Nutr J . 2015;14(1):1–7. doi: 10.1186/s12937-015-0067-3 Amro WA, Al-Qaisi W, Al-Razem F. Production and purification of IgY antibodies from chicken egg yolk. J Genet Eng Biotechnol . 2018;16(1):99–103. doi: 10.1016/j.jgeb.2017.10.003 Koppelman SJ, Vlooswijk RAA, Knippels LMJ. Quantification of major peanut allergens Ara h 1 and Ara h 2 in the peanut varieties Runner, Spanish, Virginia, and Valencia, bred in different parts of the world. Allergy Eur J Allergy Clin Immunol . 2001;56(2):132–137. doi: 10.1034/j.1398-9995.2001.056002132.x Hong KS, Ki MR, Ullah HMA. Preventive effect of anti-VacA egg yolk immunoglobulin (IgY) on Helicobacter pylori-infected mice. Vaccine . 2018;36(3):371–380. doi: 10.1016/j.vaccine.2017.11.082 Usoltsev D, Sitnikova V, Kajava A, Uspenskaya M. Systematic FTIR spectroscopy study of the secondary structure changes in human serum albumin under various denaturation conditions. Biomolecules . 2019;9(8):1–17. doi: 10.3390/biom9080359 Yasui H, Takahashi K, Taki S. Near Infrared Photo-Antimicrobial Targeting Therapy for Candida albicans. Adv Ther . 2021;4(2):1–12. doi: 10.1002/adtp.202000221 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7161972","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":498178529,"identity":"65a4bb58-a971-4f7c-adfe-106b49262763","order_by":0,"name":"Kanaka Durga Devi Nelluri","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1UlEQVRIiWNgGAWjYFAC5sYDDAwSDAzsDUCOgQUxWhgbIFp4DoC0SBCtBQgkEsAkYQ3m7I0Nh3n3WNjLz3x+dcOPAgkG/vbuBLxaLHsONhzmeSaRuOF2TtnNHqDDJM6c3YBXi8GNRKCWAxIJBtI5aTd4gFoMJHIJaLn/EKwF6LAzaTf/EKXlBiNYC2PDDfZjt4mz5Uxiw8E5B4B+OZPDdlvGQIKHsF+OHz744M2BOnv59uPPbr75YyPH396LXwsS4DEAk8QqBwH2B6SoHgWjYBSMghEEAIdoTCvluXHNAAAAAElFTkSuQmCC","orcid":"","institution":"KVSR Siddhartha College of Pharmaceutical Sciences","correspondingAuthor":true,"prefix":"","firstName":"Kanaka","middleName":"Durga Devi","lastName":"Nelluri","suffix":""},{"id":498178530,"identity":"afc74e3e-fe81-4279-b1ca-b145dd255c20","order_by":1,"name":"Anupama Ammulu Manne","email":"","orcid":"","institution":"PVPSIT","correspondingAuthor":false,"prefix":"","firstName":"Anupama","middleName":"Ammulu","lastName":"Manne","suffix":""},{"id":498178531,"identity":"21ca4089-401a-4b15-8005-21c94d6028da","order_by":2,"name":"Praveen Kumar Vemuri","email":"","orcid":"","institution":"Koneru Lakshmaiah Education Foundation","correspondingAuthor":false,"prefix":"","firstName":"Praveen","middleName":"Kumar","lastName":"Vemuri","suffix":""},{"id":498178532,"identity":"724905da-0682-4be6-bbb5-9efbd5a3e42a","order_by":3,"name":"Tanvitha Vadlamudi","email":"","orcid":"","institution":"KVSR Siddhartha College of Pharmaceutical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Tanvitha","middleName":"","lastName":"Vadlamudi","suffix":""},{"id":498178533,"identity":"60176b6b-685f-4801-8df7-2ee713169a3e","order_by":4,"name":"Prathyusha Chowdary Koduri","email":"","orcid":"","institution":"KVSR Siddhartha College of Pharmaceutical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Prathyusha","middleName":"Chowdary","lastName":"Koduri","suffix":""},{"id":498178534,"identity":"36e1f5b5-1a3a-44eb-99da-0cc8dd0b3be5","order_by":5,"name":"Vaka koti reddy","email":"","orcid":"","institution":"KVSR Siddhartha College of Pharmaceutical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Vaka","middleName":"koti","lastName":"reddy","suffix":""},{"id":498178535,"identity":"c117d441-b923-4fa8-b701-7aa4d5decc2b","order_by":6,"name":"Kallam Dhanvanth reddy","email":"","orcid":"","institution":"KVSR Siddhartha College of Pharmaceutical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Kallam","middleName":"Dhanvanth","lastName":"reddy","suffix":""}],"badges":[],"createdAt":"2025-07-19 05:23:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7161972/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7161972/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":88982075,"identity":"00e4b8d3-543d-4087-ba58-34376f953143","added_by":"auto","created_at":"2025-08-13 11:50:43","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":195812,"visible":true,"origin":"","legend":"\u003cp\u003eSDS-PAGE analysis of purified immunoglobulin Y from different eggs. \u003cstrong\u003eB\u003c/strong\u003e: IgY extracted from brown egg, \u003cstrong\u003eD\u003c/strong\u003e: IgY extracted from duck egg, \u003cstrong\u003eW\u003c/strong\u003e: IgY extracted from white egg, \u003cstrong\u003eM\u003c/strong\u003e: molecular mass marker.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7161972/v1/75f7ef0f34f0cb68600e5707.png"},{"id":88982074,"identity":"3704d3ca-7e86-421e-98f9-18f4320e01b1","added_by":"auto","created_at":"2025-08-13 11:50:43","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":65287,"visible":true,"origin":"","legend":"\u003cp\u003eSDS-PAGE analysis of peanut proteins. \u003cstrong\u003eA\u003c/strong\u003e: protein samples before dialysis, \u003cstrong\u003eB\u003c/strong\u003e: protein marker,\u003cstrong\u003e C\u003c/strong\u003e: protein samples after dialysis.\u003c/p\u003e","description":"","filename":"image2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7161972/v1/67a3a479f1866d62602a22f9.jpeg"},{"id":88982076,"identity":"96211ce6-8655-4703-a7d7-2ed6caea1cee","added_by":"auto","created_at":"2025-08-13 11:50:43","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":129870,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR peaks of peanut proteins before and after dialysis\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-7161972/v1/15bdb315ba374582a527c12b.png"},{"id":88983732,"identity":"7ca0412f-74f1-49e8-b7af-78971bb331aa","added_by":"auto","created_at":"2025-08-13 12:06:43","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":37547,"visible":true,"origin":"","legend":"\u003cp\u003eIncreasing concentration of IgY activity towards peanut antigen by ELISA\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-7161972/v1/c69db228cecf57fae5028a1a.png"},{"id":88983221,"identity":"b189f5b5-6fd9-4687-a97b-37dfa03aafd3","added_by":"auto","created_at":"2025-08-13 11:58:43","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":55678,"visible":true,"origin":"","legend":"\u003cp\u003eImmunoblot analysis of peanut extract conducted under reducing and non-reducing conditions. mAb 1: Anti-Ara h three monoclonal antibody, Biotin MACT-F0864, mAB 2: Anti-Ara h 6 (PA16H3G6) with chemiluminescent detection of 1 μg of Ara h 1 (∼64–60 kDa) and Ara h 6 (∼13–15 kDa).\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-7161972/v1/320d70ccd76e7a803bcb22b1.png"},{"id":90225974,"identity":"b54c2b36-62b3-4c2f-a088-f57a8fdd86de","added_by":"auto","created_at":"2025-08-30 11:31:45","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1374133,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7161972/v1/2c0f0395-f846-404e-b1ba-4a4e9d01ff77.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Avian immunoglobulin Y immunological response to the major peanut allergens","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe peanut (\u003cem\u003eArachis hypogaea\u003c/em\u003e L.) along with its wild cousins are native to South America, which are previously been found in Bolivia, Argentina, Paraguay, Uruguay, and present-day Brazil. An uncommon hybridization occurrence between two diploid species, \u003cem\u003eArachis duranensis\u003c/em\u003e (AA genome) and \u003cem\u003eArachis ipaensis\u003c/em\u003e (BB genome), produced the tetraploid (AABB genome) cultured peanuts that are grown today. They belong to the legume family (Fabaceae) \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Many potent allergenic proteins in peanuts can cause serious anaphylaxis reactions, even in small doses. A sizable segment of the US populace is in contact with peanuts. However, it was shown that 2.1% of children under the age of 18 and 1.4% of adults in the US generally suffer from peanut and tree nut allergies. While peanut allergy sufferers experience peanut-related anaphylaxis very frequently, fatalities are still extremely uncommon.\u003c/p\u003e\u003cp\u003eAllergen immunotherapy, which has been used for over a century and is usually given as subcutaneous injections of soluble allergen extracts, is an effective treatment for many IgE-mediated allergies \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Subcutaneous injections containing peanut extract were explored more than 20 years ago. Despite their effectiveness in treating allergies to insects and the environment, the technique was abandoned because of the high rate of anaphylaxis that followed injections \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. In the last ten years, researchers have changed the way allergen immunotherapy is administered to treat food allergies to reduce side effects while continuously exposing allergic patients to the allergen that causes their symptoms. These antigen delivery methods include ingesting the allergen orally (oral immunotherapy; OIT), administering a soluble protein extract sublingually (sublingual immunotherapy; SLIT), and applying dry allergens topically (epicutaneous immunotherapy; EPIT).\u003c/p\u003e\u003cp\u003eBlocking Fel d1's binding spots, or epitopes prevents it from binding with IgE and, thus, from inducing an allergic reaction, which is another method of easing allergy symptoms. This is just one of the ways allergen-specific immunotherapy (ASIT) works to increase the afflicted person's anti-Fel d1 IgG antibodies production. These particular IgG antibodies attach to Fel d1, blocking its IgE binding and, consequently, the allergic response \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Although ASIT has been utilized in allergy treatment and has a complex mechanism of action, ASIT has several disadvantages, including cost, reaction time, and potential side effects.\u003c/p\u003e\u003cp\u003eIt has been shown that anti-Fel d1-specific polyclonal immunoglobulin Y (sIgY) in chicken egg yolks counteracts the allergic effects of Fel d1 mast cell culture techniques \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. The avian immunoglobulin known as IgY is comparable to mammalian IgG. To give passive immunity to their young ones, hens naturally create these antibodies in response to antigen exposure. These antibodies are then transported and concentrated into egg yolks. Therefore, large amounts build up in the yolks of chicken eggs, which can be removed, refined, and used to make food. By attaching to active binding sites on the targeted proteins, these antibodies successfully lessen the antigenicity and allergenicity of those proteins. Oral administration of antibodies derived from chicken eggs has been shown in numerous studies to be both safe and effective in reducing diarrhoea in domestic animals \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eEggs naturally contain IgY antibodies, which are generally not harmful. They also produce more quickly and at a lower cost than vaccinations and may be kept in powder form at low moisture for months without refrigeration. There are other general benefits to using IgY antibodies in immunotherapy. From an animal welfare view, egg collection is non-painful and kind to the animals. Furthermore, compared to mammalian IgG, IgY has a higher binding avidity to target antigens. Because of the evolutionary difference between mammals and birds, IgY antibodies can be generated against conserved mammalian proteins more readily and effectively than IgG antibodies in other animals. Moreover, fewer antigens are required for an effective immune response \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eProtein molecules called antibodies are created in response to an antigen. They are frequently employed in research, diagnosis, and therapy because of their capacity to bind to specific targets. Most of the antibodies on the market are made by animals, particularly tiny rodents \u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. However, particular antigens elicit modest immune responses or are even entirely non-immunogenic, making generating antibodies in mammals difficult. Additionally, the process through which mammals produce antibodies necessitates the inoculation, blood sample collection, and slaughter of animals \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Egg yolk antibodies (IgY) are gaining popularity as researchers look for methods that are more effective, affordable, and refined in their usage of animals.\u003c/p\u003e\u003cp\u003eAdditionally, the percentage of particular IgY generated by hens ranges from 1 to 10% of all antibodies \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. However, the quantity of antigen administered to the hen and its immunogenicity and molecular weight are connected to the amount of antibodies generated \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. IgY technology has been employed for numerous human and veterinary health objectives due to its productive nature and more refined approach to animal welfare \u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe \u003cem\u003eArachis hypogea\u003c/em\u003e, or peanut plant, is a bean/legume family Fabaceae member \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. Even small amounts of peanut allergens can trigger symptoms, ranging from mild reactions such as itching to more severe manifestations like anaphylaxis, which is life-threatening. The peanut plant, scientifically known as \u003cem\u003eA. hypogea\u003c/em\u003e, contains several allergens that can stimulate the specific IgE antibodies production in individuals prone to peanut allergens \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Distinct populations in diverse geographic regions can have different sensitivity patterns to these allergies \u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. The major allergens are Ara h 1, Ara h 2, Ara h 3 and Ara h 6. Particularly severe allergy responses are linked to some allergens \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003ePeanuts have two bicupin seed storage proteins, Ara h 1 and Ara h 3. They belong to the highly functionally varied protein group known as the cupin superfamily\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. Peanuts also contain other seed storage proteins known as Ara h 2, Ara h 6 and Ara h 7. These proteins are seed storage proteins as of the prolamin superfamily's 2S albumin class \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. Prolamins are a collection of plant storage proteins commonly found in cereal grains such as wheat, barley and maize.\u003c/p\u003e\u003cp\u003eIn this research, the yolks of white, brown hen, and duck eggs, IgY antibodies were isolated. SDS PAGE helps to check the purity of extracted proteins and IgY antibodies. The immune activity of peanut antigen towards IgY antibodies from egg yolks was demonstrated through ELISA. Western blotting was performed to confirm antigen-antibody reaction.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cb\u003eExtraction and Purification of IgY from chicken and duck eggs\u003c/b\u003e\u003c/p\u003e\u003cp\u003eDifferent varieties of chicken eggs, such as white and brown eggs and duck eggs, are taken, and the yolk is parted from the white. The yolk is then put into a 50 ml tube, and its volume is recorded \u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. Phosphate buffer saline (PBS) is added twice the egg yolk volume, and then 3.5% polyethylene glycol (PEG 6000) of the entire volume is added and vortexed for ten minutes. The samples undergo a 20-minute, 10,000 rpm centrifugation at 4\u003csup\u003e0\u003c/sup\u003eC. After filtering the supernatant and transferring it to a new tube, 8.5% PEG 6000 (estimated based on the revised volume) is added and vortexed as previously described. Once more, the samples are centrifuged at 4 \u003csup\u003e0\u003c/sup\u003eC for 20 minutes at 10,000 rpm. The pellet is then meticulously dissolved in 1 ml of PBS with the help of a glass stick or vortexer, and the volume of PBS is increased to 10 ml. The sample is combined with 12% (w/v) PEG 6000 and vortexed similarly. Using a glass stick or vortexer, the pellet is thoroughly dissolved in 800 \u0026micro;l of PBS after the samples are centrifuged for 20 minutes at 10,000 rpm at 4 \u003csup\u003e0\u003c/sup\u003eC. After transferring the sample into a dialysis tube, rinsed it with 400 \u0026micro;l PBS and introduced it to the dialysis machine. The extract is dialyzed in 0.1% saline using a magnetic stirrer and slowly swirled all night. PBS was used in place of the saline the next day, and dialyzing was done for an additional three hours. The finished samples are examined on SDS-PAGE and kept at -20 \u003csup\u003e0\u003c/sup\u003eC \u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSDS PAGE confirmation of IgY\u003c/b\u003e\u003c/p\u003e\u003cp\u003eUsing SDS-PAGE gel electrophoresis, the isolated IgY\u0026rsquo;s molecular weight was assessed \u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. It was completed in a vertical gel slab system. Each well received 10 mL of each sample, which was then migrated for two hours at a constant 100 V. After migration, gels were dyed using Coomassie Brilliant Blue R-250.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSoluble peanut protein extraction from peanut flour\u003c/b\u003e\u003c/p\u003e\u003cp\u003eLightly roasted and finely ground peanut flour was prepared. PBS was combined with peanut flour in a 1:4 (w/v) ratio \u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. The mixture was undergone centrifugation at 15,000 rpm for 45 minutes at 4 \u003csup\u003e0\u003c/sup\u003eC after being agitated for 1.5 h with 6 M NaOH to maintain a constant pH of 8.5. The supernatant of the protein extraction was filtered through dialysis. The dialysis membrane was soaked in water for 10\u0026ndash;15 minutes before incorporating the sample. The filled Dialysis membrane was immersed in 10 mM phosphate buffer, and using the magnetic stirrer the sample was stirred for 2 h. The extract was diluted with PBS for SDS-PAGE analysis.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSDS-PAGE analysis of purified peanut proteins\u003c/b\u003e\u003c/p\u003e\u003cp\u003eA volume of 10 ul was added to the loading buffer and incubated at 100 \u003csup\u003e0\u003c/sup\u003eC for 8 minutes. When the gel loading dye reached the end of the gel, electrophoresis was conducted for 2 h at 100V with samples loaded along with a medium-range prepared protein marker. Gels remained dyed with Coomassie Brilliant Blue R-250 following migration. Destaining was done in 25% methanol, 10% acetic acid and 65% distilled water \u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003e\u003cb\u003eFTIR analysis of proteins\u003c/b\u003e\u003c/p\u003e\u003cp\u003eA Bruker Tensor 27 FTIR spectrometer (Bruker, Germany) was used to collect the spectra \u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. In a wavenumber range of 4000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e to 400 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, spectra of each sample were collected at room temperature with a resolution of 4 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. Normalization of data is done by the software named OPUS. Matlab 2014b (Mathworks, Natick MA, USA) was used by in-house software to carry out the final analysis \u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003e\u003cb\u003eImmunological activity of IgY towards peanut antigens\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTo determine the allergen's selectivity towards IgY, Affymetrix eBioscience 96-well plates coated with a 10 \u0026micro;g/ml allergen concentration and incubated in the room for two hours \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. 200 \u0026micro;l of blocking buffer was added to each well after three rounds of washing with 1X Tris-buffered saline and 0.1% Tween\u0026reg; 20 detergent (TBST) wash buffer. The wells were then left to remain at room temperature for an hour. The primary antibodies (IgY) were loaded and incubated for 30 minutes at room temperature. The mixing of a solution containing a secondary antibody (HRP conjugated Goat anti-chicken IgY antibody) (Santa-Cruz)) Conjugation to an enzyme detects the attached antibody after unbound antibodies are washed away. Unbound conjugate is removed after incubation, and 100 \u0026micro;l of the substrate solution (TMB, eBiosciences) is added. Utilizing an ELISA plate reader, substrate converted to coloured product is measured.\u003c/p\u003e\u003cp\u003e\u003cb\u003eWestern blotting\u003c/b\u003e\u003c/p\u003e\u003cp\u003eSDS-PAGE was performed using BioRad SDS-PAGE equipment in line with the directions in the GeNei Western Blotting Teaching Kit. The resultant gel was rinsed three times with distilled water at room temperature before being fixed for 15 minutes in a solution of acetic acid, isopropanol, and water (10:25:65). According to the manufacturer's suggestions, the gel was sandwiched inside the GeNei blotting cassette to transfer bands onto the nylon membrane and electrophoresed at 50V for two hours \u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThe membrane was carefully removed from the cassette and submerged in 10 ml of newly made blocking buffer for the night at 4 \u003csup\u003e0\u003c/sup\u003eC. The following day, the buffer was thrown away, and the membrane was washed thrice with 1X PBST (phosphate-buffered saline, 0.05% Tween-20) while being incubated for 3 minutes. After that, the membrane was incubated for 30 minutes at room temperature with light shaking in 10 ml of primary IgY antibody solution. After a second round of cleaning as directed, the membrane was incubated with 10 ml of horseradish peroxidase-conjugated anti-chicken IgG for 30 minutes at room temperature with light shaking. The alkaline phosphates substrate solution in 10 ml was incubated while being gently stirred for five to ten minutes to check for the intensity of the coloured band after the wash procedure was completed four times. The membrane was tested after being washed with distilled water, and the findings were noted.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cb\u003eSDS-PAGE confirmation of IgY\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFor the pure IgY of white hen eggs, SDS-PAGE analysis showed two major bands with heavy and light chains. The heavy chain with 75 kDa and the light chain with 18 kDa was confirmed through SDS-PAGE (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The rest of the two eggs (brown hen egg and duck egg) contain more IgY-like proteins, which may act as contaminants or inhibitors during antigen-antibody interactions. As white hen eggs showed the presence of pure IgY, it was chosen for further study.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eSDS-PAGE analysis of peanut proteins\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAll protein concentrations of 1 mg/ml in SDS sample buffer were adjusted before same amounts of total protein (8 \u0026micro;g) were added into each sample for the analysis. SDS-PAGE was conduceted to confirm the presence of peanut antigens in the extracted samples (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Ara h 1, Ara h 2, Ara h 3 acidic form, Ara h 3 basic form, Ara h 6 are present in peanut extracts. Ara h 3 basic structure is more prominent in after dialysis sample.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eFTIR analysis of peanut proteins\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFTIR spectroscopy of peanut antigens before and after dialysis can reveal details about the modifications in molecule structure and composition throughout the dialysis procedure. Before dialysis, the FTIR spectrum of peanut antigens exhibited distinctive absorption peaks corresponding to the numerous functional groups, including amide bonds, carbohydrates, lipids, and nucleic acids, present in the antigens. The spectrum may also point to some peanut allergens that can cause allergy reactions in sensitive people, including Ara h 1, Ara h 2, Ara h 3, and Ara h 6 (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eELISA for IgY against antigen\u003c/b\u003e\u003c/p\u003e\u003cp\u003eELISA experiment was performed using an increasing concentration of IgY antibodies with a constant concentration of peanut antigen (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The maximum absorbance was identified at 500 mg/ml of IgY antibody for all three eggs.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eWestern blotting\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe Genetix transblot device was used to blot proteins onto 0.2 \u0026micro;m nitrocellulose membranes for two hours at 125 mA (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Then the membranes were probed with a specific monoclonal antibody to epitopes on purified IgY (primary antibody) overnight at 4 \u003csup\u003e0\u003c/sup\u003eC. An enzyme-conjugated secondary antibody was coupled to the membranes for an hour following washing. Immunoblot analysis of a peanut sample probed with specific antibodies against major peanut allergens, including Ara h1, Ara h3, and Ara h6. Primary antibodies against Ara h1, Ara h3, and Ara h6 were added to the peanut extract after it had been separated by gel electrophoresis and moved on a membrane.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIgY technology is a rapidly expanding subject in the biological sciences nowadays. IgY's in body lotions, cosmetics, yoghurt, and other functional foods, as well as powder-form supplements, may be able to prevent or treat human diseases. Reducing the dosage of antibiotics used to treat bacterial infections of the digestive tract is possible with IgY antibodies. Furthermore, egg yolk antibodies offer a fresh strategy for treating intestinal parasites and \u003cem\u003eCandida albicans\u003c/em\u003e \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eSDS-PAGE was used to evaluate the fractions obtained after IgY extraction (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The projected molecular masses were matched by the molecular weight patterns for the heavy and light chains, which were 75 and 18 kDa, respectively\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. The strength of the Ara h 1 (62 kD) monomer band was noted based on the protein ladder (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In the peanut protein sample-buffer-soluble fractions of the SDS, the aggregated protein bands were more noticeable (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The outcome was consistent with a few previous research investigations \u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. As the peanuts were roasted, strong connections were found between aggregate formation seen in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and decreased in water-soluble Ara h1. This suggested that heat processes may lead to the water-soluble proteins being broken down by free radicals attacking the peptide bonds and side chains of the proteins, or they could induce the proteins to assemble by chemical cross-linking.\u003c/p\u003e\u003cp\u003eBirds have a strong immunological response to mammalian antigens because of the evolutionary separation between mammals and birds. This improves IgY's binding specificity, as demonstrated by immunological diagnostic techniques such as immunohistochemistry, ELISA, and immunofluorescence \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. This may be attributed to a loss of IgY binding because of structural changes in peanut proteins, as opposed to brown and duck eggs, where there was a low detection of IgY concentration following an increase in protein concentration (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003e). In white eggs, the concentration of IgY antibodies showed a good sensitivity with an increase in the concentration of the \u003cem\u003eArachis hypogea\u003c/em\u003e antigens. A fascinating investigation was conducted on the pro-carcinogenic \u003cem\u003eHelicobacter pylori\u003c/em\u003e infection, which affects the stomach of humans and emits vacuolating cytotoxin A (VacA), which promotes the bacteria's persistence. There have been reports that oral administration of IgY against this toxin minimizes the histological alterations generated in the stomach tissue and protects against \u003cem\u003eHelicobacter pylori\u003c/em\u003e infection \u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eSince the secondary structure of proteins is associated with the amide I band (1600\u0026ndash;1700 cm\u0026thinsp;\u0026minus;\u0026thinsp;1). It was chosen for further study because it is the most sensitive vibrational band of the protein backbone. According to previous studies, the bands that corresponded to the secondary structures \u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e. The α-helix was represented by the bonds at 1650\u0026ndash;1660 cm\u0026thinsp;\u0026minus;\u0026thinsp;1, the β-sheet by 1618\u0026ndash;1640 and 1670\u0026ndash;1690 cm\u0026thinsp;\u0026minus;\u0026thinsp;1, the β-turn by 1660\u0026ndash;1670 and 1690\u0026ndash;1700 cm\u0026thinsp;\u0026minus;\u0026thinsp;1, and the random coils by 1645 cm\u0026thinsp;\u0026minus;\u0026thinsp;1. The band areas of linked peaks were used to calculate the percentages of every single secondary structural component. The dialysis process aims to remove small molecules and impurities, so a reduction in the intensity of peaks associated with these impurities might be observed. However, the main structural features of the peanut antigens, including the characteristic absorption peaks, are not expected to be significantly altered by dialysis alone.\u003c/p\u003e\u003cp\u003ePeanut protein extract transmitted to a membrane, and probed with a specific monoclonal antibody against Ara h1, Ara h3 and Ara h6. Following primary and secondary antibody incubations, the blot was developed using chemiluminescence, and the resulting bands were analyzed for the presence and molecular weight of allergic proteins. This approach provides insights into the Ara h1\u0026rsquo;s specificity and expression in the peanut protein extract. We observed distinct banding patterns for the peanut allergens Ara h1, Ara h3, and Ara h6 when probed with specific monoclonal antibodies. The monoclonal antibody against Ara h1 revealed a prominent band at 64 KDa, consistent with the expected size of Ara h1. When probing for Ara h3, a distinct band at 48 KDa was detected, indicating the presence of Ara h3 in the peanut protein extract. Similarly, the monoclonal antibody against Ara h6 identified a specific band at 14.5 KDa, confirming the presence of Ara h6. These results suggest the co-existence of Ara h1, Ara h3, and Ara h6 in the peanut sample, providing valuable insights into the allergenic protein composition.\u003c/p\u003e\u003cp\u003eIn an in vivo skin infection model, the IgY immunophototherapy was very specific, efficacious, and did not harm the healthy epithelium. By changing only the specific antibody, this innovative method of eliminating germs specifically could be utilized for anti-infection treatment, perhaps decreasing the adversative effects of antibiotic use due to selection for resistant variations \u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003ePeanut proteins can be investigated as a significant source of products that have immunological responses. The ELISA test examined the antigenicity. The present work established that among the three eggs chosen, the concentration of IgY antibodies in the white egg shows a good sensitivity with an increase in the concentration of the peanut antigens. Western blotting was performed to confirm that this is the same protein acting as antigen or allergen during an immunological response. However, SDS-PAGE analysis of peanut proteins confirmed the presence of peanut antigens in the extracted sample. Western blotting confirmed the activity of Ara h3 towards IgY antibody. The IgY's successful performance encourages further research into its in vivo efficacy and clinical trials \u003cem\u003eagainst A. hypogea\u003c/em\u003e. Thanks to their tremendous potential, igY antibodies have a bright future in the treatment of many different diseases.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eIgG\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eimmunoglobulin G\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eIgY\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eimmunoglobulin Y\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eIgE\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eimmunoglobulin E\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eSDS PAGE\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eSodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eELISA\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eEnzyme-linked Immunosorbent assay\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003ePBS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003ePhosphate buffer saline\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003ePEG 6000\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003epolyethylene glycol\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eTBST\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003e1X Tris-buffered saline with 0.1% Tween\u0026reg; 20 detergent\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003e1X PBST\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003ephosphate-buffered saline\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003e0.05% Tween-20.\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003c/div\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e-Ethics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eI/We\u0026nbsp;Kanaka Durga Devi.N,\u0026nbsp;Anupama .A. Manne, Praveen Kumar Vemuri, Tanvitha Vadlamudi, Prathyusha Chowdary Koduri, Vaka koti reddy, Kallm Dhanvanth reddy, the author(s) of manuscript entitled \u0026nbsp;“\u003cstrong\u003eAvian immunoglobulin Y immunological response to the major peanut allergens\u003c/strong\u003e” do hereby authorize you to publish the above said manuscript in the journal. \u0026nbsp;I/We further state that:\u003c/p\u003e\n\u003col start=\"1\" type=\"1\"\u003e\n \u003cli\u003eThis article/Paper authored by me/us is an original and genuine work. It does not infringe on the right of others and does not contain any libelous or unlawful statement. It has neither been submitted for review/publication nor published elsewhere in any print/electronic form. The tables, figures or data are not copied from anyother publications.\u003c/li\u003e\n \u003cli\u003e\u0026nbsp;I/We permit authorities concerned to publish the said paper in the journal with editorial modifications, if any.\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003e\u003cstrong\u003e-Consent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eI/We\u0026nbsp;Kanaka Durga Devi.N,\u0026nbsp;Anupama .A. Manne, Praveen Kumar Vemuri, Tanvitha Vadlamudi, Prathyusha Chowdary Koduri, Vaka koti reddy, Kallm Dhanvanth reddy, the author(s) of manuscript entitled \u0026nbsp;“\u003cstrong\u003eAvian immunoglobulin Y immunological response to the major peanut allergens\u003c/strong\u003e” do hereby authorize you to publish the above said manuscript in the journal\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e-Availability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e-Competing interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors do not have any conflict of interest”.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e-Funding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e-Authors' contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed to the study conception and design. Material preparation, project administration, supervision, first draft of the manuscript was written by \u003cstrong\u003eManne Anupama Ammulu\u003c/strong\u003e. Data collection, analysis, methodology, manuscript review and editing by \u003cstrong\u003eKanaka durga devi Nelluri.\u003c/strong\u003e Visualization and analysis by\u003cstrong\u003e\u0026nbsp;Praveen Kumar Vemuri\u003c/strong\u003e. Data collection and methodology, drafting manuscript by\u003cstrong\u003eTanvitha Vadlamudi, Prathyusha Chowdary Koduri, Vaka koti reddy, Kallm Dhanvanth reddy\u003c/strong\u003e. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e-Acknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors are thankful to the Principal and management of KVSR Siddhartha college of pharmaceutical sciences, Vijayawada for providing all the facilities for this research project.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eZhuang W, Chen H, Yang M. The genome of cultivated peanut provides insight into legume karyotypes, polyploid evolution and crop domestication. \u003cem\u003eNat Genet\u003c/em\u003e. 2019;51(5):865\u0026ndash;876. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41588-019-0402-2\u003c/span\u003e\u003cspan address=\"10.1038/s41588-019-0402-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eViswanathan RK, Busse WW. Allergen immunotherapy in allergic respiratory diseases: From mechanisms to meta-analyses. \u003cem\u003eChest\u003c/em\u003e. 2012;141(5):1303\u0026ndash;1314. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1378/chest.11-2800\u003c/span\u003e\u003cspan address=\"10.1378/chest.11-2800\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJohnson-Weaver BT, Staats HF, Burks AW, Kulis MD. Adjuvanted immunotherapy approaches for peanut allergy. \u003cem\u003eFront Immunol\u003c/em\u003e. 2018;9(SEP):1\u0026ndash;8. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fimmu.2018.02156\u003c/span\u003e\u003cspan address=\"10.3389/fimmu.2018.02156\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSatyaraj E, Sun P, Sherrill S. Fel d1 Blocking Antibodies: A Novel Method to Reduce IgE-Mediated Allergy to Cats. \u003cem\u003eJ Immunol Res\u003c/em\u003e. 2021;2021. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1155/2021/5545173\u003c/span\u003e\u003cspan address=\"10.1155/2021/5545173\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSatyaraj E, Li Q, Sun P, Sherrill S. Anti-Fel d1 immunoglobulin Y antibody-containing egg ingredient lowers allergen levels in cat saliva. \u003cem\u003eJ Feline Med Surg\u003c/em\u003e. 2019;21(10):875\u0026ndash;881. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1177/1098612X19861218\u003c/span\u003e\u003cspan address=\"10.1177/1098612X19861218\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAbbas AT, El-Kafrawy SA, Sohrab SS, Azhar EIA. IgY antibodies for the immunoprophylaxis and therapy of respiratory infections. \u003cem\u003eHum Vaccines Immunother\u003c/em\u003e. 2019;15(1):264\u0026ndash;275. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/21645515.2018.1514224\u003c/span\u003e\u003cspan address=\"10.1080/21645515.2018.1514224\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eA. Michael1,, S. Meenatchisundaram2* GP, Subbraj1 T, Selvakumaran3 R, Ramalingam4 and S. Chicken egg yolk antibodies (IgY) as an alternative to mammalian antibodies. \u003cem\u003eIndian J Sci Technol\u003c/em\u003e. 2010;6(1):7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://journals.sagepub.com/doi/10.1177/1120700020921110%0Ahttps://doi.org/10.1016/j.reuma.2018.06.001%0Ahttps://doi.org/10.1016/j.arth.2018.03.044%0Ahttps://reader.elsevier.com/reader/sd/pii/S1063458420300078?token=C039B\u003c/span\u003e\u003cspan address=\"http://journals.sagepub.com/doi/10.1177/1120700020921110%0Ahttps://doi.org/10.1016/j.reuma.2018.06.001%0Ahttps://doi.org/10.1016/j.arth.2018.03.044%0Ahttps://reader.elsevier.com/reader/sd/pii/S1063458420300078?token=C039B\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e8B13922A2079230DC9AF11A333E295FCD8\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePauly D, Chacana PA, Calzado EG, Brembs B, Schade R. Igy technology: Extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation. \u003cem\u003eJ Vis Exp\u003c/em\u003e. 2011;(51):1\u0026ndash;6. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3791/3084\u003c/span\u003e\u003cspan address=\"10.3791/3084\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLarsson A, Campbell A, Eriksson M. Chicken antibodies are highly suitable for particle enhanced turbidimetric assays. \u003cem\u003eFront Immunol\u003c/em\u003e. 2022;13(October):1\u0026ndash;8. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fimmu.2022.1016781\u003c/span\u003e\u003cspan address=\"10.3389/fimmu.2022.1016781\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAgurto-Arteaga A, Poma-Acevedo A, Rios-Matos D. Preclinical Assessment of IgY Antibodies Against Recombinant SARS-CoV-2 RBD Protein for Prophylaxis and Post-Infection Treatment of COVID-19. \u003cem\u003eFront Immunol\u003c/em\u003e. 2022;13(May):1\u0026ndash;12. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fimmu.2022.881604\u003c/span\u003e\u003cspan address=\"10.3389/fimmu.2022.881604\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePereira EPV, van Tilburg MF, Florean EOPT, Guedes MIF. Egg yolk antibodies (IgY)and their applications in human and veterinary health: A review. \u003cem\u003eInt Immunopharmacol\u003c/em\u003e. 2019;73(May):293\u0026ndash;303. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.intimp.2019.05.015\u003c/span\u003e\u003cspan address=\"10.1016/j.intimp.2019.05.015\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eIqbal A, Shah F, Hamayun M. Allergens of Arachis hypogaeaand the effect of processing on their detection by ELISA. \u003cem\u003eFood Nutr Res\u003c/em\u003e. 2016;60(February). doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3402/fnr.v60.28945\u003c/span\u003e\u003cspan address=\"10.3402/fnr.v60.28945\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePalladino C, Breiteneder H. Peanut allergens. \u003cem\u003eMol Immunol\u003c/em\u003e. 2018;100(March):58\u0026ndash;70. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.molimm.2018.04.005\u003c/span\u003e\u003cspan address=\"10.1016/j.molimm.2018.04.005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVereda A, Van Hage M, Ahlstedt S. Peanut allergy: Clinical and immunologic differences among patients from 3 different geographic regions. \u003cem\u003eJ Allergy Clin Immunol\u003c/em\u003e. 2011;127(3):603\u0026ndash;607. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jaci.2010.09.010\u003c/span\u003e\u003cspan address=\"10.1016/j.jaci.2010.09.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStorni F, Zeltins A, Balke I. Vaccine against peanut allergy based on engineered virus-like particles displaying single major peanut allergens. \u003cem\u003eJ Allergy Clin Immunol\u003c/em\u003e. 2020;145(4):1240\u0026ndash;1253.e3. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jaci.2019.12.007\u003c/span\u003e\u003cspan address=\"10.1016/j.jaci.2019.12.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDunwell JM, Purvis A, Khuri S. Cupins: The most functionally diverse protein superfamily? \u003cem\u003ePhytochemistry\u003c/em\u003e. 2004;65(1):7\u0026ndash;17. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.phytochem.2003.08.016\u003c/span\u003e\u003cspan address=\"10.1016/j.phytochem.2003.08.016\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMylne JS, Hara-Nishimura I, Rosengren KJ. Seed storage albumins: Biosynthesis, trafficking and structures. \u003cem\u003eFunct Plant Biol\u003c/em\u003e. 2014;41(7):671\u0026ndash;677. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1071/FP14035\u003c/span\u003e\u003cspan address=\"10.1071/FP14035\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePauly D, Chacana PA, Calzado EG, Brembs B, Schade R. Igy technology: Extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation. \u003cem\u003eJ Vis Exp\u003c/em\u003e. 2011;i(51):2\u0026ndash;7. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3791/3084\u003c/span\u003e\u003cspan address=\"10.3791/3084\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAKITA EM, NAKAI S. Immunoglobulins from Egg Yolk: Isolation and Purification. \u003cem\u003eJ Food Sci\u003c/em\u003e. 1992;57(3):629\u0026ndash;634. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1365-2621.1992.tb08058.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1365-2621.1992.tb08058.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKirley TL, Norman AB. Unfolding of IgG domains detected by non-reducing SDS-PAGE. \u003cem\u003eBiochem Biophys Res Commun\u003c/em\u003e. 2018;503(2):944\u0026ndash;949. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bbrc.2018.06.100\u003c/span\u003e\u003cspan address=\"10.1016/j.bbrc.2018.06.100\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBerglund JP, Szczepanski N, Penumarti A. Preparation and Analysis of Peanut Flour Used in Oral Immunotherapy Clinical Trials. \u003cem\u003eJ Allergy Clin Immunol Pract\u003c/em\u003e. 2017;5(4):1098\u0026ndash;1104. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jaip.2016.11.034\u003c/span\u003e\u003cspan address=\"10.1016/j.jaip.2016.11.034\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNowakowski AB, Wobig WJ, Petering DH. Native SDS-PAGE: High resolution electrophoretic separation of proteins with retention of native properties including bound metal ions. \u003cem\u003eMetallomics\u003c/em\u003e. 2014;6(5):1068\u0026ndash;1078. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1039/c4mt00033a\u003c/span\u003e\u003cspan address=\"10.1039/c4mt00033a\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMiller LM, Bourassa MW, Smith RJ. FTIR spectroscopic imaging of protein aggregation in living cells. \u003cem\u003eBiochim Biophys Acta - Biomembr\u003c/em\u003e. 2013;1828(10):2339\u0026ndash;2346. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bbamem.2013.01.014\u003c/span\u003e\u003cspan address=\"10.1016/j.bbamem.2013.01.014\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStrych U, Willson RC. Purification and Characterization of Proteins. \u003cem\u003eMan Ind Microbiol Biotechnol\u003c/em\u003e. 2014;10(4):731\u0026ndash;742. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1128/9781555816827.ch52\u003c/span\u003e\u003cspan address=\"10.1128/9781555816827.ch52\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHnasko RM, Lin A V., McGarvey JA, Mattison CP. Sensitive and selective detection of peanut allergen Ara h 1 by ELISA and lateral flow immunoassay. \u003cem\u003eFood Chem\u003c/em\u003e. 2022;396(July):133657. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.foodchem.2022.133657\u003c/span\u003e\u003cspan address=\"10.1016/j.foodchem.2022.133657\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMishra M, Tiwari S, Gomes A V. Protein purification and analysis: Next generation western blotting techniques. \u003cem\u003eExpert Rev Proteomics\u003c/em\u003e. 2017;14(11):1037\u0026ndash;1053. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/14789450.2017.1388167\u003c/span\u003e\u003cspan address=\"10.1080/14789450.2017.1388167\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eM\u0026uuml;ller S, Schubert A, Zajac J, Dyck T, Oelkrug C. IgY antibodies in human nutrition for disease prevention. \u003cem\u003eNutr J\u003c/em\u003e. 2015;14(1):1\u0026ndash;7. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s12937-015-0067-3\u003c/span\u003e\u003cspan address=\"10.1186/s12937-015-0067-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAmro WA, Al-Qaisi W, Al-Razem F. Production and purification of IgY antibodies from chicken egg yolk. \u003cem\u003eJ Genet Eng Biotechnol\u003c/em\u003e. 2018;16(1):99\u0026ndash;103. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jgeb.2017.10.003\u003c/span\u003e\u003cspan address=\"10.1016/j.jgeb.2017.10.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKoppelman SJ, Vlooswijk RAA, Knippels LMJ. Quantification of major peanut allergens Ara h 1 and Ara h 2 in the peanut varieties Runner, Spanish, Virginia, and Valencia, bred in different parts of the world. \u003cem\u003eAllergy Eur J Allergy Clin Immunol\u003c/em\u003e. 2001;56(2):132\u0026ndash;137. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1034/j.1398-9995.2001.056002132.x\u003c/span\u003e\u003cspan address=\"10.1034/j.1398-9995.2001.056002132.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHong KS, Ki MR, Ullah HMA. Preventive effect of anti-VacA egg yolk immunoglobulin (IgY) on Helicobacter pylori-infected mice. \u003cem\u003eVaccine\u003c/em\u003e. 2018;36(3):371\u0026ndash;380. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.vaccine.2017.11.082\u003c/span\u003e\u003cspan address=\"10.1016/j.vaccine.2017.11.082\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eUsoltsev D, Sitnikova V, Kajava A, Uspenskaya M. Systematic FTIR spectroscopy study of the secondary structure changes in human serum albumin under various denaturation conditions. \u003cem\u003eBiomolecules\u003c/em\u003e. 2019;9(8):1\u0026ndash;17. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/biom9080359\u003c/span\u003e\u003cspan address=\"10.3390/biom9080359\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYasui H, Takahashi K, Taki S. Near Infrared Photo-Antimicrobial Targeting Therapy for Candida albicans. \u003cem\u003eAdv Ther\u003c/em\u003e. 2021;4(2):1\u0026ndash;12. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/adtp.202000221\u003c/span\u003e\u003cspan address=\"10.1002/adtp.202000221\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Arachis hypogea, ELISA, FTIR, IgY, SDS PAGE, Western blotting","lastPublishedDoi":"10.21203/rs.3.rs-7161972/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7161972/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eIntroduction\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSpecifically in animal models of human infectious illnesses, chicken immunoglobulin Y (IgY), a highly conserved counterpart of human immunoglobulin G (IgG), has demonstrated advantages and a favourable safety profile. IgY is inexpensive, quick-acting, and simple to make. Egg-laying chickens can easily produce huge quantities of IgY antibodies without damaging the environment or infrastructural expenditure. Over 30% of people suffer from immunoglobulin E (IgE)-mediated allergy, which is the most prevalent hypersensitivity illness. Althoughabout 1%-2% of the general population suffers from peanut allergy, peanuts are one of the most pervasive food causes of deadly anaphylaxis globally.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eArachis hypogea\u003c/em\u003econtains allergens that exhibit immunological responses towards IgY antibodies. Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis \u0026nbsp;(SDS PAGE) analysis was performed to check the purity of extracted peanut and IgY samples, acquiring the molecular weight patterns of heavy and light chains of 75 and 18 kDa. The peanut proteins were screened for their activity against immunoglobulins for allergic responses through the Enzyme-linked Immunosorbent assay (ELISA) technique. Among the white hen, brown hen and duck eggs, the white eggs' concentration of IgY antibodies showed an excellent sensitivity with increased attention to the \u003cem\u003eArachis hypogea\u003c/em\u003e antigens.FTIR analysis of peanut proteins was carried out to identify extracted proteins. Antigen-antibody interaction was confirmed through immunoblotting. We observed distinct banding patterns for the peanut allergens Ara h1, Ara h3, and Ara h6 when probed with specific monoclonal antibodies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDesigning therapeutic IgY antibodies that can passively suppress IgE-mediated reactions in allergy patients is another potential future approach.\u003c/p\u003e","manuscriptTitle":"Avian immunoglobulin Y immunological response to the major peanut allergens","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-13 11:50:38","doi":"10.21203/rs.3.rs-7161972/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a40d63ee-c7cf-41ea-b9b4-a418758614c6","owner":[],"postedDate":"August 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-08-30T11:23:37+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-13 11:50:38","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7161972","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7161972","identity":"rs-7161972","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}