Imperial College London
Alternate

Professor Sergei Kazarian

Faculty of Engineering, Department of Chemical Engineering

Professor of Physical Chemistry
 
 
 
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Contact

 

+44 (0)20 7594 5574s.kazarian Website

 
 
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Assistant

 

Mrs Sarah Payne +44 (0)20 7594 5567

 
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Location

 

440Bone BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Beattie:2021:10.1039/D1AN00985K,
author = {Beattie, J and Rowland-Jones, R and Farys, M and Kucia-Tran, R and Kazarian, S and Byrne, B},
doi = {10.1039/D1AN00985K},
journal = {The Analyst},
pages = {5177--5185},
title = {Insight into purification of monoclonal antibodies in industrial columns via studies of Protein A binding capacity by in situ ATR-FTIR spectroscopy},
url = {http://dx.doi.org/10.1039/D1AN00985K},
volume = {146},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Therapeutic monoclonal antibodies (mAbs) are effective treatments for a range of cancers and other serious diseases, however mAb treatments cost on average ∼$100000 per year per patient, limiting their use. Currently, industry favours Protein A affinity chromatography (PrAc) as the key step in downstream processing of mAbs. This step, although highly efficient, represents a significant mAb production cost. Fouling of the Protein A column and Protein A ligand leaching contribute to the cost of mAb production by shortening the life span of the resin. In this study, we assessed the performance of used PrAc resin recovered from the middle inlet, center and outlet as well as the side inlet of a pilot-scale industrial column. We used a combination of static binding capacity (SBC) analysis and Attenuated Total Reflection-Fourier Transform Infrared (ATR-FTIR) spectroscopy to explore the used resin samples. SBC analysis demonstrated that resin from the inlet of the column had lower binding capacity than resin from the column outlet. ATR-FTIR spectroscopy with PLS (partial least square) analysis confirmed the results obtained from SBC analysis. Importantly, in situ ATR-FTIR spectroscopy also allowed both measurement of the concentration and assessment of the conformational state of the bound Protein A. Our results reveal that PrAc resin degradation after use is dependent on column location and that neither Protein A ligand leaching nor denaturation are responsible for binding capacity loss.
AU  - Beattie,J
AU  - Rowland-Jones,R
AU  - Farys,M
AU  - Kucia-Tran,R
AU  - Kazarian,S
AU  - Byrne,B
DO  - 10.1039/D1AN00985K
EP  - 5185
PY  - 2021///
SN  - 0003-2654
SP  - 5177
TI  - Insight into purification of monoclonal antibodies in industrial columns via studies of Protein A binding capacity by in situ ATR-FTIR spectroscopy
T2  - The Analyst
UR  - http://dx.doi.org/10.1039/D1AN00985K
UR  - https://pubs.rsc.org/en/content/articlelanding/2021/AN/D1AN00985K
UR  - http://hdl.handle.net/10044/1/90682
VL  - 146
ER  -
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