Generation, Diversity Determination, and Application to Antibody Selection of a Human Naive Fab Library.

ABSTRACT: We constructed a large naïve human Fab library (3 × 10¹⁰ colonies) from the lymphocytes of 809 human donors, assessed available diversities of the heavy-chain variable (VH) and κ light-chain variable (VK) domain repertoires, and validated the library by selecting Fabs against 10 therapeutically relevant antigens by phage display. We obtained a database of unique 7,373 VH and 41,804 VK sequences by 454 pyrosequencing, and analyzed the repertoires. The distribution of VH and VK subfamilies and germline genes in our antibody repertoires slightly differed from those in earlier published natural antibody libraries. The frequency of somatic hypermutations (SHMs) in heavy-chain complementarity determining region (HCDR)1 and HCDR2 are higher compared with the natural IgM repertoire. Ana

SUBMITTER: Kim S 

PROVIDER: S-EPMC5638773 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature