Isolation of a nanobody specific to the PstS-1 protein and evaluation of its immunoreactivity with structural components of Mycobacterium tuberculosis granuloma
preprint
OA: closed
Abstract
M. tuberculosis (Mtb) causes infectious granulomatous disease tuberculosis (TB). Within TB granuloma, foci of Mtb secreted antigens anchored on the surface of either bacilli or host cells may serve as targetable biomarkers for antibody based molecular imaging of TB. Nanobody is better suited over conventional antibody or its fragment derivatives for molecular imaging due to its quick localization in target tissue and rapid clearance from off-target organs. Here, we report the production of a high affinity nanobody against PstS-1 protein of Mtb which helps bacilli in phosphate uptake as well as host cell adhesion. C8 nanobody was isolated from a phage display library displaying nanobodies which was constructed from a camel immunized with secretory proteins of Mtb. C8 Nb was characterized in vitro and in vivo for its immunoreactivity against PstS-1 protein. Ability of C8 Nb to bind PstS-1 protein present on the surface of Mtb bacilli or adhered on macrophages and its localization around BCG cells injected intramuscularly in mice demonstrate its potential in the development of molecular imaging for tuberculosis.
Full text
2,048 characters
· extracted from
oa-doi-fallback
· click to expand
Abstract
M. tuberculosis (Mtb) causes infectious granulomatous disease tuberculosis (TB). Within TB granuloma, foci of Mtb secreted antigens anchored on the surface of either bacilli or host cells may serve as targetable biomarkers for antibody based molecular imaging of TB. Nanobody is better suited over conventional antibody or its fragment derivatives for molecular imaging due to its quick localization in target tissue and rapid clearance from off-target organs. Here, we report the production of a high affinity nanobody against PstS-1 protein of Mtb which helps bacilli in phosphate uptake as well as host cell adhesion. C8 nanobody was isolated from a phage display library displaying nanobodies which was constructed from a camel immunized with secretory proteins of Mtb. C8 Nb was characterized in vitro and in vivo for its immunoreactivity against PstS-1 protein. Ability of C8 Nb to bind PstS-1 protein present on the surface of Mtb bacilli or adhered on macrophages and its localization around BCG cells injected intramuscularly in mice demonstrate its potential in the development of molecular imaging for tuberculosis.
Abbreviations
- Nb
- Nanobody
- VHH
- Variable domain of the heavy chain of the heavy-chain only antibody
- VH
- Variable domain of the heavy chain of the conventional antibody
- IgG
- Immunoglobulin G
- SDS-PAGE
- Sodium dodecyl sulfate polyacrylamide gel electrophoresis
- Fabs
- Antigen-binding fragments
- PBMCs
- Peripheral blood mononuclear cells
- PFU
- Plaque-forming units
- BSA
- Bovine serum albumin
- E. coli
- Escherichia coli
- HRP
- Horseradish peroxidase
- ELISA
- Enzyme-linked immunosorbent assay
- SPR
- Surface plasmon resonance
- IPTG
- Isopropyl-β-d-thiogalactoside
- 6xHis
- Hexahistidine
- pH
- Potential of hydrogen
- mAbs
- Monoclonal antibodies
- FBS
- Fetal bovine serum
- KD
- Equilibrium dissociation constant
- Ka
- Association rate constant
- Kd
- Dissociation rate constant
- OD600
- Optical density at 600nm wavelength
- ADC
- Albumin dextrose catalase
- CPM
- Counts per minute
Text is read by the "Ask this paper" AI Q&A widget below. Extraction quality varies by source — PMC NXML preserves structure cleanly, OA-HTML may include some navigation residue, and OA-PDF can have broken hyphenation. The publisher copy (via DOI) is the canonical version.
My notes (saved in your browser only)
Ask this paper
Answers must be backed by verbatim quotes from this paper's full text. Hallucinated quotes are dropped automatically; if no verbatim passage answers the question, we say so. How this works
Citation neighborhood (no data yet)
We don't have any in-corpus citations linked to this paper yet. This is a recent paper (2025) — citers typically take a year or two to land, and the OpenAlex reference graph may still be filling in.
Source provenance
- europepmc
- last seen: 2026-05-20T01:45:00.602351+00:00