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10.1080/19420862.2018.1426422

http://scihub22266oqcxt.onion/10.1080/19420862.2018.1426422
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suck abstract from ncbi


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pmid29376776      MAbs 2018 ; 10 (3): 431-43
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  • A natively paired antibody library yields drug leads with higher sensitivity and specificity than a randomly paired antibody library #MMPMID29376776
  • Adler AS; Bedinger D; Adams MS; Asensio MA; Edgar RC; Leong R; Leong J; Mizrahi RA; Spindler MJ; Bandi SR; Huang H; Tawde P; Brams P; Johnson DS
  • MAbs 2018[Apr]; 10 (3): 431-43 PMID29376776show ga
  • Deep sequencing and single-chain variable fragment (scFv) yeast display methods are becoming more popular for discovery of therapeutic antibody candidates in mouse B cell repertoires. In this study, we compare a deep sequencing and scFv display method that retains native heavy and light chain pairing with a related method that randomly pairs heavy and light chain. We performed the studies in a humanized mouse, using interleukin 21 receptor (IL-21R) as a test immunogen. We identified 44 high-affinity binder scFv with the native pairing method and 100 high-affinity binder scFv with the random pairing method. 30% of the natively paired scFv binders were also discovered with the randomly paired method, and 13% of the randomly paired binders were also discovered with the natively paired method. Additionally, 33% of the scFv binders discovered only in the randomly paired library were initially present in the natively paired pre-sort library. Thus, a significant proportion of ?randomly paired? scFv were actually natively paired. We synthesized and produced 46 of the candidates as full-length antibodies and subjected them to a panel of binding assays to characterize their therapeutic potential. 87% of the antibodies were verified as binding IL-21R by at least one assay. We found that antibodies with native light chains were more likely to bind IL-21R than antibodies with non-native light chains, suggesting a higher false positive rate for antibodies from the randomly paired library. Additionally, the randomly paired method failed to identify nearly half of the true natively paired binders, suggesting a higher false negative rate. We conclude that natively paired libraries have critical advantages in sensitivity and specificity for antibody discovery programs.
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