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Construction of a Human Antibody Domain (VH) Library

  • Weizao Chen
  • Zhongyu Zhu
  • Xiaodong Xiao
  • Dimiter S. Dimitrov
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 525)

Abstract

Highly diverse antibody (Fab or scFv) libraries have become vital sources to select antibodies with high affinity and novel properties. Combinatorial strategies provide efficient ways of creating antibody libraries containing a large number of individual clones. These strategies include the reassembly of naturally occurring genes encoding the heavy and light chains from either immune or nonimmune B-cell sources, or introduction of synthetic diversity to either the framework regions (FRs) or the complementarity-determining regions (CDRs) of the variable domains of antibodies. In the late 1980s, the smallest known antigen-binding fragment was identified when a murine VH repertoire was screened for binding to lysozyme. This fragment (∼15 kDa), called a “domain antibody”, or “dAb”, is approximately four times smaller than a Fab and half the size of a scFv. Here, we describe the construction of a phage-displayed VH library and an approach to introduce genetic diversity in this library, where both diverse human CDRs and synthetic CDRs are combined into a single-domain (VH) framework.

Key words

Library construction phage display domain antibody human VH CDRs grafting diversity 

Notes

Acknowledgments

This project was supported by the NIH Intramural AIDS Targeted Antiviral Program (IATAP) and the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Weizao Chen
    • 1
  • Zhongyu Zhu
    • 1
  • Xiaodong Xiao
    • 1
  • Dimiter S. Dimitrov
    • 1
  1. 1.NCI-Frederick, National Institutes of HealthFrederickUSA

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