Yeast mating for combinatorial Fab library generation and surface display.
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.
Weaver-Feldhaus
Jane M
JM
Pacific Northwest National Laboratory, MSIN:K4-12, 902 Battelle Boulevard, P.O. Box 999, Richland, WA 99352, USA.
Lou
Jianlong
J
Coleman
James R
JR
Siegel
Robert W
RW
Marks
James D
JD
Feldhaus
Michael J
MJ
D000915:Antibody Affinity; D000918:Antibody Specificity; D020650:Combinatorial Chemistry Techniques; D005822:Genetic Vectors; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000276:immunology; D019151:Peptide Library; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015003:Yeasts / Q000235:genetics*
D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library