Escherichia coli inner membrane display system for high-throughput screening of dimeric proteins

Migyeong Jo, Bora Hwang, Hyun Woung Yoon, Sang Taek Jung

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)2849-2858
Number of pages10
JournalBiotechnology and Bioengineering
Volume115
Issue number12
DOIs
Publication statusPublished - 2018 Dec 1
Externally publishedYes

Fingerprint

Escherichia coli
Screening
Display devices
Throughput
Proteins
Membranes
Membrane Proteins
Antibodies
Polypeptides
Antigens
Immunoglobulin G
Cytokines
Light
Peptides
Enzymes

Keywords

  • antibody
  • bacterial display
  • directed evolution
  • library screening
  • multimeric protein

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

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

In: Biotechnology and Bioengineering, Vol. 115, No. 12, 01.12.2018, p. 2849-2858.

Research output: Contribution to journalArticle

@article{6feaaf9c47aa4e6d8cac34f1e4306795,
title = "Escherichia coli inner membrane display system for high-throughput screening of dimeric proteins",
abstract = "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.",
keywords = "antibody, bacterial display, directed evolution, library screening, multimeric protein",
author = "Migyeong Jo and Bora Hwang and Yoon, {Hyun Woung} and Jung, {Sang Taek}",
year = "2018",
month = "12",
day = "1",
doi = "10.1002/bit.26826",
language = "English",
volume = "115",
pages = "2849--2858",
journal = "Biotechnology and Bioengineering",
issn = "0006-3592",
publisher = "Wiley-VCH Verlag",
number = "12",

}

TY - JOUR

T1 - Escherichia coli inner membrane display system for high-throughput screening of dimeric proteins

AU - Jo, Migyeong

AU - Hwang, Bora

AU - Yoon, Hyun Woung

AU - Jung, Sang Taek

PY - 2018/12/1

Y1 - 2018/12/1

N2 - 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.

AB - 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.

KW - antibody

KW - bacterial display

KW - directed evolution

KW - library screening

KW - multimeric protein

UR - http://www.scopus.com/inward/record.url?scp=85053769723&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85053769723&partnerID=8YFLogxK

U2 - 10.1002/bit.26826

DO - 10.1002/bit.26826

M3 - Article

C2 - 30171695

AN - SCOPUS:85053769723

VL - 115

SP - 2849

EP - 2858

JO - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

SN - 0006-3592

IS - 12

ER -