Multimer formation is indispensable to the intrinsicbiologicalfunctions of many natural proteins. For example, the human immunoglobulin G (IgG) antibody has two variable regions (heavy chain variable domain [VH] and light chain variable domain [VL]) that must be assembled for specific antigen binding, and homodimerization of the antibody's Fc domain is essential for eliciting therapeutic effector functions. For the more efficient high‐throughput directed evolution of multimeric proteins with ease of cultivation and handling, here we report a membrane protein drift and assembly (MPDA) system, in which a multimeric protein is displayed on a bacterial inner membrane by drifting and auto‐assembling membrane‐anchored subunit polypeptides. This system enabled the auto‐assembly of membrane‐tethered Fv domains (VH and VL) or the monomeric Fc domain into a functional hetero‐ or homodimeric protein complex on the bacterial inner membrane. This system could also be used to enrich a desired engineered Fc variant from a mixture containing a million‐fold excess of wild‐type Fc domain, indicating the applicability of the MPDA system for the high‐throughput directed evolution of a variety of multimeric proteins, such as cytokines, enzymes, or structural proteins.

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membranetethered fv domains monomeric fc domain intrinsicbiologicalfunctions cytokines enzymes engineered fc variant drifting and autoassembling membraneanchored subunit hetero or homodimeric protein chain variable domain vh igg antibody highthroughput directed evolution of multimeric proteins therapeutic effector membrane protein drift and assembly mpda system structural proteins immunoglobulin antibodys fc domain

Migyeong Jo, Bora Hwang, Hyun Woung Yoon, Sang Taek Jung,.Escherichia coli inner membrane display system for high‐throughput screening of dimeric proteins. 115 (115),2849-2858.

Migyeong Jo, Bora Hwang, Hyun Woung Yoon, Sang Taek Jung,"Escherichia coli inner membrane display system for high‐throughput screening of dimeric proteins" 115

Migyeong Jo, Bora Hwang, Hyun Woung Yoon, Sang Taek Jung,"Escherichia coli inner membrane display system for high‐throughput screening of dimeric proteins"