Journal
FEBS LETTERS
Volume 564, Issue 1-2, Pages 24-34Publisher
WILEY
DOI: 10.1016/S0014-5793(04)00309-6
Keywords
Fab; yeast display; selection; library; antibody; single chain Fv
Funding
- NCI NIH HHS [U54 CA90788] Funding Source: Medline
- NIAID NIH HHS [U01 AI056493, R21 AI53389-01] Funding Source: Medline
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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.
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