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Construction of a scFv Library with Synthetic, Non-combinatorial CDR Diversity

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1575)

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.

Key words

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

Notes

Acknowledgment

This work was supported by the National Research Foundation of Korea (NRF) grant for Medical Bioconvergence Research Center (NRF-2013M3A6A4044991) and the Bio & Medical Technology Development Program of the NRF funded by the Korean government, MSIP (NRF-2015M3A9B6029138) to H.S.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Life ScienceEwha Womans UniversitySeoulKorea
  2. 2.Department of Bioinspired ScienceEwha Womans UniversitySeoulKorea

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