Computational design and immunoinformatic validation of a multistage mRNA vaccine candidate against Mycobacterium tuberculosis

Computational design and immunoinformatic validation of a multistage mRNA vaccine candidate against Mycobacterium tuberculosis | 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 Computational design and immunoinformatic validation of a multistage mRNA vaccine candidate against Mycobacterium tuberculosis Manika sharma, Dr. Parul Bhatt, Dr. Medha Singh, Dr. Kiran Bharat Lokhande, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9037759/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 15 You are reading this latest preprint version Abstract Tuberculosis (TB),the biggest cause of death from any known infectious disease, has been a problem for the world's health system for many years. The only approved vaccine is BCG and now a number of vaccines are undergoing clinical trials. The newly recognised mRNA vaccines can provide a good alternative to the traditional vaccine. Therefore, the goal of this work is to use computational techniques to build a multi-stage tuberculosis mRNA vaccine. Nine multistage-expressing Mycobacterium tuberculosis (Mtb) proteins that have been linked to cell entry, pathogenesis, and dormancy regulon were used. Eighteen promiscuous, proinflammatory, non-toxic antigenic epitopes that bound to both MHC classes (I and II) were chosen from nine proteins. Co-translational structural components (5'm7G cap, UTRs, Kozak sequence, Poly A tail) were incorporated into the design of the mRNA vaccine, which was predicted to be extremely stable. Additionally, molecular docking reveals the vaccine candidate’s interaction with TLR2 and TLR4 immunological receptors by establishing a Leucine-Rich-Repeats (LRR) specific interface. Molecular dynamics (MD) simulations were also performed to evaluate the structural stability and dynamical behaviour of the vaccine candidate in complex with TLR2 and TLR4. Following the simulations, binding free energy calculations were conducted using the Molecular Mechanics/Poisson–Boltzmann Surface Area (MM/PBSA) method, which indicated stable thermodynamic binding of the vaccine candidate with both the receptors. Immune simulation tests using C-Immsim confirmed that the translated vaccine construct was immunogenic. Overall, it was anticipated that this multi-stage expressing mRNA vaccine would be highly immunogenic, stable, safe, and antigenic. Our method for creating an immunoinformatic-based mRNA vaccine can be an attractive tactic seems promising against tuberculosis, however, experimental validation needs to be carried out. Mycobacterium tuberculosis immuno-informatics mRNA vaccine peptide-based vaccine candidates. Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementarydata.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 08 Apr, 2026 Reviews received at journal 08 Apr, 2026 Reviews received at journal 07 Apr, 2026 Reviews received at journal 06 Apr, 2026 Reviewers agreed at journal 25 Mar, 2026 Reviews received at journal 24 Mar, 2026 Reviewers agreed at journal 24 Mar, 2026 Reviewers agreed at journal 24 Mar, 2026 Reviewers agreed at journal 24 Mar, 2026 Reviewers agreed at journal 24 Mar, 2026 Reviewers agreed at journal 23 Mar, 2026 Reviewers invited by journal 23 Mar, 2026 Editor assigned by journal 06 Mar, 2026 Submission checks completed at journal 06 Mar, 2026 First submitted to journal 05 Mar, 2026 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. 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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-9037759","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":618038194,"identity":"fdb39437-1265-4906-80a2-d144c889dfab","order_by":0,"name":"Manika sharma","email":"","orcid":"","institution":"University of Delhi","correspondingAuthor":false,"prefix":"","firstName":"Manika","middleName":"","lastName":"sharma","suffix":""},{"id":618038195,"identity":"a6c98c7e-f800-4a85-9c75-4e12993c4b8a","order_by":1,"name":"Dr. Parul Bhatt","email":"","orcid":"","institution":"University of 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