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J Mol Biol. 2002 Aug 2;321(1):49-56.

Phage versus phagemid libraries for generation of human monoclonal antibodies.

Author information

1
Department of Anesthesia and Pharmaceutical Chemistry, University of California, San Francisco, Rm 3C-38, San Francisco General Hospital, 1001 Potrero Ave, 94110, USA.

Abstract

Non-immune (naïve) phage antibody libraries have become an important source of antibodies for reagent, diagnostic, and therapeutic use. To date, reported naïve libraries have been constructed in phagemid vectors as fusions to pIII, yielding primarily single copy (monovalent) display of antibody fragments. For this work, we subcloned the single chain Fv (scFv) gene repertoire from a naïve phagemid antibody library into a true phage vector to create a multivalently displayed scFv phage library. Compared to monovalently displayed scFv, multivalent phage display resulted in improved efficiency of display as well as antibody selection. A greater number of antibodies were obtained and at earlier rounds of selection. Such increased efficiency allows the screening for binding antibodies after a single round of selection, greatly facilitating automation. Expression levels of antigen-binding scFv were also higher than from the phagemid library. In contrast, the affinities of scFv from the phage library were lower than from the phagemid library. This could be overcome by utilizing the scFv in a multivalent format, by affinity maturation, or by converting the library to monovalent display after the first round of selection.

PMID:
12139932
DOI:
10.1016/s0022-2836(02)00561-2
[Indexed for MEDLINE]

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