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Cell-free selection of domain antibodies by in vitro compartmentalization.

Authors
  • Sepp, Armin1
  • Griffiths, Andrew
  • 1 Innovation Biopharm Discovery Unit, Biopharm R&D, GlaxoSmithKline Plc, Cambridge, UK. [email protected]
Type
Published Article
Journal
Methods in molecular biology (Clifton, N.J.)
Publication Date
Jan 01, 2012
Volume
911
Pages
183–198
Identifiers
DOI: 10.1007/978-1-61779-968-6_12
PMID: 22886253
Source
Medline
License
Unknown

Abstract

Efficient identification of antibodies, or any fragments thereof, displaying desired specificity and affinity is critical for the development of novel immunotherapeutics. Here we describe the adaptation of in vitro compartmentalization for the cell-free selection of Vκ and VH domain antibodies (dAbs™) from large combinatorial libraries. The dAbs™ are in vitro expressed in fusion to the N-terminus of single-chain variant of phage P22 Arc repressor DNA-binding domain that links the compartmentally expressed protein molecules to their encoding PCR fragment-based genes via cognate operator sites present on the DNA. Libraries of up to 10(10) in size can be rapidly assembled and selected for improved affinity in equilibrium and off-rate conditions.

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