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

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• 作者：Song Lin ; Nga Rewa Houston-Cummings ; Bianka Prinz ; René ; e Moore ; Beata Bobrowicz ; Robert C. Davidson ; Stefan Wildt ; Terrance A. Stadheim ; Dongxing Zha ; ^{dongxing_zha@merck.com}
• 关键词：Surface display ; Glycoengineered Pichia ; Fragment of antigen binding (Fab) ; Antibody expression ; Antibody affinity ; FACS
• 刊名：Journal of Immunological Methods
• 出版年：2012
• 出版时间：31 January 2012
• 年：2012
• 卷：375
• 期：1-2
• 页码：159-165
• 全文大小：701 K

文摘

A fragment 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 Man₅GlcNAc₂ 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.