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J Mol Biol. 2018 Oct 19;430(21):4387-4400. doi: 10.1016/j.jmb.2018.09.003. Epub 2018 Sep 11.

A Sortase A Programmable Phage Display Format for Improved Panning of Fab Antibody Libraries.

Author information

1
Department of Immunology and Microbiology, The Scripps Research Institute, Jupiter, FL 33458, USA.
2
Department of Immunology and Microbiology, The Scripps Research Institute, Jupiter, FL 33458, USA. Electronic address: crader@scripps.edu.

Abstract

Phage display of combinatorial antibody libraries is a versatile tool in the field of antibody engineering, with diverse applications including monoclonal antibody (mAb) discovery, affinity maturation, and humanization. To improve the selection efficiency of antibody libraries, we developed a new phagemid display system that addresses the complication of bald phage propagation. The phagemid facilitates the biotinylation of fragment of antigen binding (Fab) antibody fragments displayed on phage via Sortase A catalysis and the subsequent enrichment of Fab-displaying phage during selections. In multiple contexts, this selection approach improved the enrichment of target-reactive mAbs by depleting background phage. Panels of cancer cell line-reactive mAbs with high diversity and specificity were isolated from a naïve chimeric rabbit/human Fab library using this approach, highlighting its potential to accelerate antibody engineering efforts and to empower concerted antibody drug and target discovery.

KEYWORDS:

antibody engineering; phagemid; site-specific programming; sortagging; whole-cell panning

PMID:
30213726
PMCID:
PMC6186506
[Available on 2019-10-19]
DOI:
10.1016/j.jmb.2018.09.003

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