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

Neutron reflectivity measurement of protein A-antibody complex at the solid-liquid interface

Mazzer, AR; Clifton, LA; Perevozchikova, T; Butler, PD; Roberts, CJ; Bracewell, DG; (2017) Neutron reflectivity measurement of protein A-antibody complex at the solid-liquid interface. Journal of Chromatography A , 1499 pp. 118-131. 10.1016/j.chroma.2017.03.084. Green open access

[thumbnail of Published article]
Preview
Text (Published article)
Mazzer_Neutron_reflectivity_measurement.pdf

Download (3MB) | Preview
[thumbnail of Supplementary Figure]
Preview
Text (Supplementary Figure)
Mazzer_Neutron_reflectivity_measurement_Supplementary.pdf

Download (314kB) | Preview

Abstract

Chromatography is a ubiquitous unit operation in the purification of biopharmaceuticals yet few studies have addressed the biophysical characterisation of proteins at the solution-resin interface. Chromatography and other adsorption and desorption processes have been shown to induce protein aggregation which is undesirable in biopharmaceutical products. In order to advance understanding of how adsorption processes might impact protein stability, neutron reflectivity was used to characterise the structure of adsorbed immunoglobulin G (IgG) on model surfaces. In the first model system, IgG was adsorbed directly to silica and demonstrated a side-on orientation with high surface contact. A maximum dimension of 60Å in the surface normal direction and high density surface coverage were observed under pH 4.1 conditions. In chromatography buffers, pH was found to influence IgG packing density and orientation at the solid-liquid interface. In the second model system, which was designed to mimic an affinity chromatography surface, protein A was attached to a silica surface to produce a configuration representative of a porous glass chromatography resin. Interfacial structure was probed during sequential stages from ligand attachment, through to IgG binding and elution. Adsorbed IgG structures extended up to 250Å away from the surface and showed dependence on surface blocking strategies. The data was suggestive of two IgG molecules bound to protein A with a somewhat skewed orientation and close proximity to the silica surface. The findings provide insight into the orientation of adsorbed antibody structures under conditions encountered during chromatographic separations.

Type: Article
Title: Neutron reflectivity measurement of protein A-antibody complex at the solid-liquid interface
Location: Netherlands
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.chroma.2017.03.084
Publisher version: https://doi.org/10.1016/j.chroma.2017.03.084
Language: English
Additional information: Copyright © 2017 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license(http://creativecommons.org/licenses/by/4.0/).
Keywords: Adsorption, Antibody, Chromatography, Interface, Neutron reflectivity, Protein A
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/1554450
Downloads since deposit
152Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item