title: Construction of a scFv Library with Synthetic, Non-combinatorial CDR Diversity.
abstract: Many large synthetic antibody libraries have been designed, constructed, and successfully generated high-quality antibodies suitable for various demanding applications. While synthetic antibody libraries have many advantages such as optimized framework sequences and a broader sequence landscape than natural antibodies, their sequence diversities typically are generated by random combinatorial synthetic processes which cause the incorporation of many undesired CDR sequences. Here, we describe the construction of a synthetic scFv library using oligonucleotide mixtures that contain predefined, non-combinatorially synthesized CDR sequences. Each CDR is first inserted to a master scFv framework sequence and the resulting single-CDR libraries are subjected to a round of proofread panning. The proofread CDR sequences are assembled to produce the final scFv library with six diversified CDRs.
authors.affiliation: Department of Life Science, Ewha Womans University, 52 Ewhayeodae-gail, Seodaemun-gu, Seoul, Korea.
authors.forename: Xuelian
authors.initials: X
authors.lastname: Bai
authors.affiliation: Department of Life Science, Ewha Womans University, 52 Ewhayeodae-gail, Seodaemun-gu, Seoul, Korea. hshim@ewha.ac.kr.|Department of Bioinspired Science, Ewha Womans University, Seoul, Korea. hshim@ewha.ac.kr.
authors.forename: Hyunbo
authors.initials: H
authors.lastname: Shim
mesh_terms: D000595:Amino Acid Sequence; D022801:Complementarity Determining Regions / Q000235:genetics* / Q000276:immunology; D015723:Gene Library; D005813:Genes, Synthetic; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology
chemical_list: D022801:Complementarity Determining Regions; D019151:Peptide Library; D057127:Single-Chain Antibodies
keywords: Antibody library; Non-combinatorial CDR diversity; Phage display; Synthetic CDR diversity; Synthetic antibody library; scFv