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