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A suspension cell‐based interaction platform for interrogation of membrane proteins
Author(s) -
Krohl Patrick J.,
Kim Kook Bum,
Lew Lance,
VanDyke Derek,
Ludwig Seth D.,
Spangler Jamie B.
Publication year - 2020
Publication title -
aiche journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.958
H-Index - 167
eISSN - 1547-5905
pISSN - 0001-1541
DOI - 10.1002/aic.16995
Subject(s) - computational biology , biopanning , cell sorting , yeast , suspension culture , cell , interrogation , microbiology and biotechnology , cell culture , recombinant dna , chemistry , biology , peptide library , biochemistry , gene , peptide sequence , genetics , archaeology , history
Abstract The majority of clinically approved therapeutics target membrane proteins (MPs), highlighting the need for tools to study this important category of proteins. To overcome limitations with recombinant MP expression, whole cell screening techniques have been developed that present MPs in their native conformations. Whereas many such platforms utilize adherent cells, here we introduce a novel suspension cell‐based platform termed “biofloating” that enables quantitative analysis of interactions between proteins displayed on yeast and MPs expressed on mammalian cells, without need for genetic fusions. We characterize and optimize biofloating and illustrate its sensitivity advantage compared to an adherent cell‐based platform (biopanning). We further demonstrate the utility of suspension cell‐based approaches by iterating rounds of magnetic‐activated cell sorting selections against MP‐expressing mammalian cells to enrich for a specific binder within a yeast‐displayed antibody library. Overall, biofloating represents a promising new technology that can be readily integrated into protein discovery and development workflows.