Production of F(ab′)2 from Monoclonal and Polyclonal Antibodies

Shai Rosenstein; Anna Vaisman-Mentesh; Limor Levy; Aya Kigel; Yael Dror; Yariv Wine

doi:10.1002/cpmb.119

Production of F(ab′)₂ from Monoclonal and Polyclonal Antibodies

Shai Rosenstein, Anna Vaisman-Mentesh, Limor Levy, Aya Kigel, Yael Dror, Yariv Wine

Tel Aviv University

Research output: Contribution to journal › Article › peer-review

Abstract

Antibodies are widely used in therapeutic, diagnostic, and research applications, and antibody derivatives such as F(ab′)₂ fragments are used when only a particular antibody region is required. F(ab′)₂ can be produced through antibody engineering, but some applications require F(ab′)₂ produced from an original formulated antibody or directly from a polyclonal antibody pool. The cysteine protease immunoglobulin-degrading enzyme (IdeS) from Streptococcus pyogenes digests immunoglobulin G (IgG) specifically and efficiently to produce F(ab′)₂. Here we detail the production and purification of recombinant IdeS; its utilization to digest monoclonal or polyclonal antibodies to F(ab′)₂ fragments; and F(ab′)₂ purification through consecutive affinity chromatography steps. The resultant F(ab′)₂ exhibit high purity, retain antigen-binding functionality, and are readily utilizable in various downstream applications.

Original language	English
Article number	e119
Journal	Current Protocols in Molecular Biology
Volume	131
Issue number	1
DOIs	https://doi.org/10.1002/cpmb.119
State	Published - 1 Jun 2020

Keywords

IdeS
mAbs
monoclonal antibody
polyclonal antibodies

All Science Journal Classification (ASJC) codes

Molecular Biology

Access to Document

10.1002/cpmb.119

Cite this

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abstract = "Antibodies are widely used in therapeutic, diagnostic, and research applications, and antibody derivatives such as F(ab′)2 fragments are used when only a particular antibody region is required. F(ab′)2 can be produced through antibody engineering, but some applications require F(ab′)2 produced from an original formulated antibody or directly from a polyclonal antibody pool. The cysteine protease immunoglobulin-degrading enzyme (IdeS) from Streptococcus pyogenes digests immunoglobulin G (IgG) specifically and efficiently to produce F(ab′)2. Here we detail the production and purification of recombinant IdeS; its utilization to digest monoclonal or polyclonal antibodies to F(ab′)2 fragments; and F(ab′)2 purification through consecutive affinity chromatography steps. The resultant F(ab′)2 exhibit high purity, retain antigen-binding functionality, and are readily utilizable in various downstream applications.",

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AU - Dror, Yael

AU - Wine, Yariv

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