Protein and Antibody Engineering by Phage Display

J. C. Frei, Jonathan R. Lai

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Phage display is an in vitro selection technique that allows for the rapid isolation of proteins with desired properties including increased affinity, specificity, stability, and new enzymatic activity. The power of phage display relies on the phenotype-to-genotype linkage of the protein of interest displayed on the phage surface with the encoding DNA packaged within the phage particle, which allows for selective enrichment of library pools and high-throughput screening of resulting clones. As an in vitro method, the conditions of the binding selection can be tightly controlled. Due to the high-throughput nature, rapidity, and ease of use, phage display is an excellent technological platform for engineering antibody or proteins with enhanced properties. Here, we describe methods for synthesis, selection, and screening of phage libraries with particular emphasis on designing humanizing antibody libraries and combinatorial scanning mutagenesis libraries. We conclude with a brief section on troubleshooting for all stages of the phage display process.

Original languageEnglish (US)
JournalMethods in Enzymology
DOIs
StateAccepted/In press - 2016

Fingerprint

Protein Engineering
Bacteriophages
Display devices
Antibodies
Proteins
Libraries
Screening
Throughput
Antibodies, Monoclonal, Humanized
Mutagenesis
Clone Cells
Genotype
Scanning
Phenotype
DNA

Keywords

  • Antibody engineering
  • Combinatorial biochemistry
  • Combinatorial scanning mutagenesis
  • Phage display
  • Protein engineering

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Protein and Antibody Engineering by Phage Display. / Frei, J. C.; Lai, Jonathan R.

In: Methods in Enzymology, 2016.

Research output: Contribution to journalArticle

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