Imperial College London
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ProfessorBernadetteByrne

Faculty of Natural Sciences, Department of Life Sciences

Associate Dean (Equality, Diversity and Inclusion) for FoNS
 
 
 
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Contact

 

+44 (0)20 7594 3004b.byrne Website

 
 
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Location

 

504Sir Ernst Chain BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Beattie:2022:10.1021/acs.analchem.2c03063,
author = {Beattie, JW and Istrate, A and Lu, A and Marshall, C and Rowland-Jones, RC and Farys, M and Kazarian, SG and Byrne, B},
doi = {10.1021/acs.analchem.2c03063},
journal = {Analytical Chemistry},
pages = {15703--15710},
title = {Causes of industrial protein a column degradation, explored using Raman spectroscopy.},
url = {http://dx.doi.org/10.1021/acs.analchem.2c03063},
volume = {94},
year = {2022}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Monoclonal antibodies (mAbs) are used extensively as biotherapeutics for chronic and acute conditions. Production of mAbs is lengthy and expensive, with protein A affinity capture the most costly step, due both to the nature of the resin and its marked reduction in binding capacity with repeated use. Our previous studies using in situ ATR-FTIR spectroscopy indicated that loss in protein A binding capacity is not the result of leaching or degradation of protein A ligand, suggesting fouling is the principal cause. Here we explore binding behavior and resin capacity loss using Raman spectroscopy. Our data reveal a distinct Raman spectral fingerprint for mAb bound to the protein A ligand of MabSelect SuRe. The results show that the drop in static binding capacity (SBC) previously observed for used protein A resin is discernible by Raman spectroscopy in combination with partial least-squares regression. The SBC is lowest (35.76 mg mL-1) for used inlet resin compared to used outlet (40.17 mg mL-1) and unused resin samples (70.35 mg mL-1). Depth profiling by Raman spectroscopy indicates that at below saturating concentrations (∼18 mg mL-1), binding of mAb is not homogeneous through used resin beads with protein binding preferentially to the outer regions of the bead, in contrast to fully homogeneous distribution through unused control MabSelect SuRe resin beads. Analysis of the Raman spectra indicates that one foulant is irreversibly bound mAb. The presence of irreversibly bound mAb and host cell proteins was confirmed by mass spectrometric analysis of used resin beads.
AU  - Beattie,JW
AU  - Istrate,A
AU  - Lu,A
AU  - Marshall,C
AU  - Rowland-Jones,RC
AU  - Farys,M
AU  - Kazarian,SG
AU  - Byrne,B
DO  - 10.1021/acs.analchem.2c03063
EP  - 15710
PY  - 2022///
SN  - 0003-2700
SP  - 15703
TI  - Causes of industrial protein a column degradation, explored using Raman spectroscopy.
T2  - Analytical Chemistry
UR  - http://dx.doi.org/10.1021/acs.analchem.2c03063
UR  - https://www.ncbi.nlm.nih.gov/pubmed/36318727
UR  - https://pubs.acs.org/doi/10.1021/acs.analchem.2c03063
UR  - http://hdl.handle.net/10044/1/100530
VL  - 94
ER  -
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