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A novel fragment of antigen binding (Fab) surface display platform using glycoengineered Pichia pastoris
Journal article   Peer reviewed

A novel fragment of antigen binding (Fab) surface display platform using glycoengineered Pichia pastoris

Song Lin, Nga Rewa Houston-Cummings, Bianka Prinz, Renee Moore, Beata Bobrowicz, Robert C. Davidson, Stefan Wildt, Terrance A. Stadheim and Dongxing Zha
Journal of immunological methods, v 375(1-2), 159
31 Jan 2012
PMID: 22019510

Abstract

Biochemical Research Methods Biochemistry & Molecular Biology Immunology Life Sciences & Biomedicine Science & Technology
Afragment of antigen binding (Fab) surface display system was developed using a glycoengineered Pichia pastoris host strain genetically modified to secrete glycoproteins with mammalian mannose-type Man(5)GlcNAc(2) N-linked glycans. The surface display method described here takes advantage of a pair of coiled-coil peptides as the linker while using the Saccharomyces cerevisiae Sed1p GPI-anchored cell surface protein as an anchoring domain. Several Fabs were successfully displayed on the cell surface using this system and the expression level of the displayed Fabs was correlated to that of secreted Fabs from the same glycoengineered host in the absence of the cell wall anchor. Strains displaying different model Fabs were mixed and, through cell sorting, the strain displaying more expressed Fab molecule or the strain displaying the Fab with higher affinity for an antigen was effectively enriched by FACS. This novel yeast surface display system provides a general platform for the display of Fab libraries for affinity and/or expression maturation using glycoengineered Pichia. (C) 2011 Elsevier B.V. All rights reserved.

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Biochemical Research Methods
Immunology
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