Yeast mating for combinatorial Fab library generation and surface display

Journal Article · Fri Apr 23 04:00:00 EDT 2004 · FEBS letters

DOI:https://doi.org/10.1016/S0014-5793(04)00309-6· OSTI ID:15020859

Feldhaus, Jane M; Lou, Jianlong; Coleman, James R; Siegel, Robert W; Marks, James D; Feldhaus, Michael

Yeast display of antibody fragments has proven to be an efficient and productive means for directed evolution of single chain Fv (scFv) 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.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 15020859
• Report Number(s): PNNL-SA-40675; 6504; NN2004000
• Journal Information: FEBS letters, Journal Name: FEBS letters Journal Issue: 1-2 Vol. 564
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
AFFINITY
ANTIBODIES
ANTIGENS
MATING
SACCHAROMYCES CEREVISIAE
STABILITY
STRAINS
VECTORS
YEASTS
environmental molecular sciences laboratory