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

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

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


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 languageEnglish
Article numbere119
JournalCurrent Protocols in Molecular Biology
Issue number1
StatePublished - 1 Jun 2020


FundersFunder number
Department of Molecular Biology
United States-Israel Binational Science Foundation2017359
Israel Science Foundation1282/17
Umeå Universitet


    • IdeS
    • mAbs
    • monoclonal antibody
    • polyclonal antibodies


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