Abstract
Abstract The current diagnostic tools for Tuberculosis (TB) are time consuming and not suitable for point of care devices. Antibody-based diagnostic approaches could solve this issue. We have used antibody phage-display to isolate chimeric mouse/human Fab antibodies that binds to the surface of live Mycobacterium tuberculosis (Mtb). For that purpose, mice were immunized with live or heat killed (HK) Mtb and splenic mRNA was used to generate 2 phage-display Fab libraries. Several rounds of bio panning were performed against whole cell live and HK Mtb and a substraction round was performed against non-tuberculous Mycobacteria. Of the 24 clones analyzed we found 6 Fab-expressing phages that specifically bind to live Mtb over HK or non-tuberculous Mycobacteria. Further characterization of these Fab antibodies consisted of recombinant expression and purification in E coli and selective binding to Mtb-specific subcellular fractions. This is the first study on Mtb-specific Fab antibodies that specifically bind a live and virulent Mtb strain. These molecules could represent a good platform to set up highly specific TB biomarkers.
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