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

Kanje, Sara; Venskutonytė, Raminta LU ; Scheffel, Julia LU ; Nilvebrant, Johan; Lindkvist-Petersson, Karin LU and Hober, Sophia (2018) In Journal of Molecular Biology 430(18). p.3427-3438
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... (More)

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.

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Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
antibody purification, calcium-dependent binding, Protein A, protein engineering, Z domain
in
Journal of Molecular Biology
volume
430
issue
18
pages
3427 - 3438
publisher
Elsevier
external identifiers
  • scopus:85048734234
ISSN
0022-2836
DOI
10.1016/j.jmb.2018.06.004
language
English
LU publication?
yes
id
4577a150-4af9-44b2-93ff-e1df84de1f86
date added to LUP
2018-07-02 10:50:09
date last changed
2019-02-20 11:21:20
@article{4577a150-4af9-44b2-93ff-e1df84de1f86,
  abstract     = {<p>Presented here is an engineered protein domain, based on Protein A, that displays a calcium-dependent binding to antibodies. This protein, Z<sub>Ca</sub>, 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 Z<sub>Ca</sub>–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.</p>},
  author       = {Kanje, Sara and Venskutonytė, Raminta and Scheffel, Julia and Nilvebrant, Johan and Lindkvist-Petersson, Karin and Hober, Sophia},
  issn         = {0022-2836},
  keyword      = {antibody purification,calcium-dependent binding,Protein A,protein engineering,Z domain},
  language     = {eng},
  month        = {06},
  number       = {18},
  pages        = {3427--3438},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Biology},
  title        = {Protein Engineering Allows for Mild Affinity-based Elution of Therapeutic Antibodies},
  url          = {http://dx.doi.org/10.1016/j.jmb.2018.06.004},
  volume       = {430},
  year         = {2018},
}