Yeast mating for combinatorial Fab library generation and surface display

FEBS Letters

Coleman, James R²  Feldhaus, Michael J²  Lou, Jianlong¹  Marks, James D¹  Siegel, Robert W²  Weaver-Feldhaus, Jane M² 

[1] Department of Anesthesia and Pharmaceutical Chemistry, Rm 3C-38, NH, San Francisco General Hospital, University of California San Francisco, 1001 Potrero, San Francisco, CA 94110, USA;Pacific Northwest National Laboratory, MSIN:K4-12, 902 Battelle Boulevard, P.O. Box 999, Richland, WA 99352, USA

关键词: Fab;    Yeast display;    Selection;    Library;    Antibody;    Single chain Fv;    scFv;    single chain fragment variable;    mAb;    monoclonal antibody;    SDCAA;    synthetic dextrose plus casein amino acids;    EGF;    epidermal growth factor;    VH;    variable heavy;    Vk;    variable light;   

DOI  :  10.1016/S0014-5793(04)00309-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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.

【 授权许可】

Unknown