Yeast mating for combinatorial Fab library generation and surface display

被引：66

作者：

Weaver-Feldhaus, JM

Lou, JL

Coleman, JR

Siegel, RW

Marks, JD

Feldhaus, MJ ^{[1
]}

机构：

[1] Pacific NW Natl Lab, MSIN K412,902 Battelle blvd,POB 999, Richland, WA 99352 USA

[2] Univ Calif San Francisco, San Francisco Gen Hosp, Dept Anesthesia & Pharmaceut Chem, San Francisco, CA 94110 USA

来源：

FEBS LETTERS | 2004年 / 564卷 / 1-2期

关键词：

Fab; yeast display; selection; library; antibody; single chain Fv;

D O I：

10.1016/S0014-5793(04)00309-6

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Yeast display of antibody fragments has proven to be an efficient and productive means for directed evolution of single chain Fv antibodies for increased affinity and thermal stability, and more recently for the display and screening of a non-immune library. In this paper, we describe an elegant and simple method for constructing large combinatorial Fab libraries for display on the surface of Saccharomyces cerevisiae, from modestly sized, and easily constructed, heavy and light chain libraries. To this end, we have constructed a set of yeast strains and a two vector system for heavy chain and light chain surface display of Fab fragments with free native amino termini. Through yeast mating of the haploid libraries, a very large heterodimeric immune Fab library was displayed on the diploids and high affinity antigen specific Fabs were isolated from the library. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

引用

页码：24 / 34

页数：11

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