Construction Of A Synthetic Single Domain Antibody Phage Display Library For Molecular Diagnostic Applications

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Date
2015-08
Authors
HAIRUL BAHARA, NUR HIDAYAH
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Abstract
Domain antibodies have been widely exploited as a scaffold for the generation of synthetic antibody libraries because of their relatively small size and simple folding mechanism. In this study, the generation of a highly diverse library using a single human framework (VH3-23(DP47)) and synthetic diversity introduced by randomly mutating naturally occurring within complementarity-determining regions (CDRs) CDR1,CDR2 and CDR3 of heavy chain yielded a library size of 10^9. The sequence diversity of all CDRs was determined from 28 randomly selected clones. Out of the 28 clones, 18 clones were conserved with different length of CDR3 and highly diverse amino acids residues. The quality of the library was also validated by panning against two different types of protein antigens; diseases and fluorescent proteins. Multiple unique target specific clones were obtained for most antigens. However, two monoclonal antibodies were successfully raised against Mycobacterium tuberculosis 16 kDa Hsp (Mtb 16 kDa Hsp) antigens which could potentially be used for therapeutics. In conclusion, the diverse repertoire of the naïve library can be used in the future to screen for binders against other potential antigens.
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Construction Of A Synthetic Single Domain Antibody Phage Display Library , For Molecular Diagnostic Applications
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