Use WuXi’s proprietary human naive antibody library to identify antibodies specific to each of the two splicing variants of antigen 1 named Ag 1a and Ag 1b.
Two splicing variants (Ag 1a & Ag 1b) differ by >20 different amino acids.
Hybridoma approach failed to yield antibodies selective for Ag 1b
Human naïve library was panned on the splicing variant b of the human antigen 1 (Ag 1b) in the presence of splicing variant a (Ag 1a) as a competitor. Following panning, rounds 2 and 3 outputs were screened by monoclonal phage ELISA. Round 3 output shows significant enrichment with Ag 1b specific clones with concurrent reduction of Ag 1a/Ag 1b cross-reactive clones. Sequence analysis of round 3 hits identified 6 unique clones, 2 of which also showed strong binding to cell-surface expressed Ag 1b (data not shown here).
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