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A suspension cell-based interaction platform for interrogation of membrane proteins.
  • +3
  • Patrick Krohl,
  • Kook Bum Kim,
  • Lance Lew,
  • Derek VanDyke,
  • Seth Ludwig,
  • Jamie Spangler
Patrick Krohl
Johns Hopkins University
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Kook Bum Kim
Johns Hopkins University
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Lance Lew
Johns Hopkins University
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Derek VanDyke
Johns Hopkins University
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Seth Ludwig
Johns Hopkins University
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Jamie Spangler
Johns Hopkins University
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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 (MACS) 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.

Peer review status:Published

01 Jun 2020Submitted to AIChE Journal
03 Jun 2020Submission Checks Completed
03 Jun 2020Assigned to Editor
05 Jun 2020Reviewer(s) Assigned
02 Jul 2020Editorial Decision: Revise Minor
20 Jul 20201st Revision Received
20 Jul 2020Submission Checks Completed
20 Jul 2020Assigned to Editor
22 Jul 2020Editorial Decision: Accept
11 Aug 2020Published in AIChE Journal. 10.1002/aic.16995