UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Protein A chromatography increases monoclonal antibody aggregation rate during subsequent low pH virus inactivation hold

Mazzer, AR; Perraud, X; Halley, J; O'Hara, J; Bracewell, DG; (2015) Protein A chromatography increases monoclonal antibody aggregation rate during subsequent low pH virus inactivation hold. Journal of Chromatography A , 1415 pp. 83-90. 10.1016/j.chroma.2015.08.068. Green open access

[thumbnail of 1-s2.0-S0021967315012650-main.pdf]
Preview
Text
1-s2.0-S0021967315012650-main.pdf

Download (1MB) | Preview

Abstract

Protein A chromatography is a near-ubiquitous method of mAb capture in bioprocesses. The use of low pH buffer for elution from protein A is known to contribute to product aggregation. Yet, a more limited set of evidence suggests that low pH may not be the sole cause of aggregation in protein A chromatography, rather, other facets of the process may contribute significantly. This paper presents a well-defined method for investigating this problem. An IgG4 was incubated in elution buffer after protein A chromatography (typical of the viral inactivation hold) and the quantity of monomer in neutralised samples was determined by size exclusion chromatography; elution buffers of different pH values predetermined to induce aggregation of the IgG4 were used. Rate constants for monomer decay over time were determined by fitting exponential decay functions to the data. Similar experiments were implemented in the absence of a chromatography step, i.e. IgG4 aggregation at low pH. Rate constants for aggregation after protein A chromatography were considerably higher than those from low pH exposure alone; a distinct shift in aggregation rates was apparent across the pH range tested.

Type: Article
Title: Protein A chromatography increases monoclonal antibody aggregation rate during subsequent low pH virus inactivation hold
Location: Netherlands
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.chroma.2015.08.068
Publisher version: http://dx.doi.org/10.1016/j.chroma.2015.08.068
Additional information: © 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Keywords: Affinity adsorption, Aggregation, IgG, Size exclusion chromatography, Unfolding pH
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/1473390
Downloads since deposit
163Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item