TY - CHAP
T1 - Protein and Antibody Engineering by Phage Display
AU - Frei, J. C.
AU - Lai, J. R.
N1 - Publisher Copyright:
© 2016 Elsevier Inc.
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
KW - Antibody engineering
KW - Combinatorial biochemistry
KW - Combinatorial scanning mutagenesis
KW - Phage display
KW - Protein engineering
UR - http://www.scopus.com/inward/record.url?scp=84977636423&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84977636423&partnerID=8YFLogxK
U2 - 10.1016/bs.mie.2016.05.005
DO - 10.1016/bs.mie.2016.05.005
M3 - Chapter
C2 - 27586328
AN - SCOPUS:84977636423
T3 - Methods in Enzymology
SP - 45
EP - 87
BT - Methods in Enzymology
PB - Academic Press Inc.
ER -