Protocol

Single Domain Antibodies

Volume 911 of the series Methods in Molecular Biology pp 431-443

Date:

Affinity Maturation of Single-Domain Antibodies by Yeast Surface Display

  • Akiko KoideAffiliated withDepartment of Biochemistry and Molecular Biology, The University of Chicago
  • , Shohei KoideAffiliated withDepartment of Biochemistry and Molecular Biology, The University of Chicago Email author 

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Abstract

Although single-domain antibodies derived from libraries prepared either after animal immunization or naïve animals generally exhibit reasonable affinity, it is often desirable to further improve their affinity. This chapter describes protocols for improving the affinity of single-domain antibodies using quantitative library sorting by yeast surface display. An example is included where we also exploit a complementary strength of phage display in generating larger sequence diversity prior to library sorting with yeast surface display.

Key words

Antibody engineering Molecular recognition Combinatorial libraries Phage display