Construction of a scFv library with synthetic, non-combinatorial CDR diversity

Xuelian Bai, Hyunbo Shim

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

2 Scopus citations

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages15-29
Number of pages15
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1575
ISSN (Print)1064-3745

Keywords

  • Antibody library
  • Non-combinatorial CDR diversity
  • Phage display
  • scFv
  • Synthetic antibody library
  • Synthetic CDR diversity

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