Protein Engineering Allows for Mild Affinity-based Elution of Therapeutic Antibodies

Research output: Contribution to journalArticle

Abstract

Presented here is an engineered protein domain, based on Protein A, that displays a calcium-dependent binding to antibodies. This protein, ZCa, is shown to efficiently function as an affinity ligand for mild purification of antibodies through elution with ethylenediaminetetraacetic acid. Antibodies are commonly used tools in the area of biological sciences and as therapeutics, and the most commonly used approach for antibody purification is based on Protein A using acidic elution. Although this affinity-based method is robust and efficient, the requirement for low pH elution can be detrimental to the protein being purified. By introducing a calcium-binding loop in the Protein A-derived Z domain, it has been re-engineered to provide efficient antibody purification under mild conditions. Through comprehensive analyses of the domain as well as the ZCa–Fc complex, the features of this domain are well understood. This novel protein domain provides a very valuable tool for effective and gentle antibody and Fc-fusion protein purification.

Details

Authors
Organisations
External organisations
  • KTH Royal Institute of Technology
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biochemistry and Molecular Biology

Keywords

  • antibody purification, calcium-dependent binding, Protein A, protein engineering, Z domain
Original languageEnglish
Pages (from-to)3427-3438
JournalJournal of Molecular Biology
Volume430
Issue number18
Early online date2018 Jun 7
Publication statusPublished - 2018
Publication categoryResearch
Peer-reviewedYes