Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody TherapeuticsCitation formats

  • External authors:
  • Bethan Mcavan
  • Leo A Bowsher
  • Thomas Powell
  • John F O'Hara
  • Mariangela Spitali
  • Royston Goodacre

Standard

Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody Therapeutics. / Mcavan, Bethan; Bowsher, Leo A; Powell, Thomas; O'Hara, John F; Spitali, Mariangela; Goodacre, Royston; Doig, Andrew.

In: Analytical Chemistry, 01.07.2020.

Research output: Contribution to journalArticlepeer-review

Harvard

Mcavan, B, Bowsher, LA, Powell, T, O'Hara, JF, Spitali, M, Goodacre, R & Doig, A 2020, 'Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody Therapeutics', Analytical Chemistry.

APA

Mcavan, B., Bowsher, L. A., Powell, T., O'Hara, J. F., Spitali, M., Goodacre, R., & Doig, A. (Accepted/In press). Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody Therapeutics. Analytical Chemistry.

Vancouver

Mcavan B, Bowsher LA, Powell T, O'Hara JF, Spitali M, Goodacre R et al. Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody Therapeutics. Analytical Chemistry. 2020 Jul 1.

Author

Mcavan, Bethan ; Bowsher, Leo A ; Powell, Thomas ; O'Hara, John F ; Spitali, Mariangela ; Goodacre, Royston ; Doig, Andrew. / Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody Therapeutics. In: Analytical Chemistry. 2020.

Bibtex

@article{1a15990789f34d27a5129777f5deb72d,
title = "Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody Therapeutics",
abstract = "Monoclonal antibodies (mAbs) represent a rapidly expanding market for biotherapeutics. Structural changes in the mAb can lead to unwanted immunogenicity, reduced efficacy and loss of material during production. The pharmaceutical sector requires new protein characterisation tools that are fast, applicable in situ and to the manufacturing process. Raman has been highlighted as a technique to suit this application as it is information rich, minimally invasive, insensitive to water background and requires little to no sample preparation. This study investigates the applicability of Raman to detect Post-Translational Modifications (PTMs) and degradation seen in mAbs. IgG4 molecules have been incubated under a range of conditions known to result in degradation of the therapeutic including varied pH, temperature, agitation, photo and chemical stresses. Aggregation was measured using size-exclusion chromatography and PTM levels were calculated using peptide mapping. By combining principal component analysis (PCA) with Raman spectroscopy and circular dichroism (CD) spectroscopy structural analysis we were able to separate proteins based on PTMs and degradation. Furthermore, by identifying key bands that lead to the PCA separation we could correlate spectral peaks to specific PTMs. In particular, we have identified a peak which exhibits a shift in samples with higher levels of Trp oxidation. Through separation of IgG4 aggregates, by size, we have shown a linear correlation between peak wavenumbers of specific functional groups and amount of aggregate present. We therefore demonstrate the capability for Raman spectroscopy to be used as an analytical tool to measure degradation and PTMs in-line with therapeutic production.",
author = "Bethan Mcavan and Bowsher, {Leo A} and Thomas Powell and O'Hara, {John F} and Mariangela Spitali and Royston Goodacre and Andrew Doig",
year = "2020",
month = jul,
day = "1",
language = "English",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",

}

RIS

TY - JOUR

T1 - Raman Spectroscopy to Monitor Post-Translational Modifications and Degradation in Monoclonal Antibody Therapeutics

AU - Mcavan, Bethan

AU - Bowsher, Leo A

AU - Powell, Thomas

AU - O'Hara, John F

AU - Spitali, Mariangela

AU - Goodacre, Royston

AU - Doig, Andrew

PY - 2020/7/1

Y1 - 2020/7/1

N2 - Monoclonal antibodies (mAbs) represent a rapidly expanding market for biotherapeutics. Structural changes in the mAb can lead to unwanted immunogenicity, reduced efficacy and loss of material during production. The pharmaceutical sector requires new protein characterisation tools that are fast, applicable in situ and to the manufacturing process. Raman has been highlighted as a technique to suit this application as it is information rich, minimally invasive, insensitive to water background and requires little to no sample preparation. This study investigates the applicability of Raman to detect Post-Translational Modifications (PTMs) and degradation seen in mAbs. IgG4 molecules have been incubated under a range of conditions known to result in degradation of the therapeutic including varied pH, temperature, agitation, photo and chemical stresses. Aggregation was measured using size-exclusion chromatography and PTM levels were calculated using peptide mapping. By combining principal component analysis (PCA) with Raman spectroscopy and circular dichroism (CD) spectroscopy structural analysis we were able to separate proteins based on PTMs and degradation. Furthermore, by identifying key bands that lead to the PCA separation we could correlate spectral peaks to specific PTMs. In particular, we have identified a peak which exhibits a shift in samples with higher levels of Trp oxidation. Through separation of IgG4 aggregates, by size, we have shown a linear correlation between peak wavenumbers of specific functional groups and amount of aggregate present. We therefore demonstrate the capability for Raman spectroscopy to be used as an analytical tool to measure degradation and PTMs in-line with therapeutic production.

AB - Monoclonal antibodies (mAbs) represent a rapidly expanding market for biotherapeutics. Structural changes in the mAb can lead to unwanted immunogenicity, reduced efficacy and loss of material during production. The pharmaceutical sector requires new protein characterisation tools that are fast, applicable in situ and to the manufacturing process. Raman has been highlighted as a technique to suit this application as it is information rich, minimally invasive, insensitive to water background and requires little to no sample preparation. This study investigates the applicability of Raman to detect Post-Translational Modifications (PTMs) and degradation seen in mAbs. IgG4 molecules have been incubated under a range of conditions known to result in degradation of the therapeutic including varied pH, temperature, agitation, photo and chemical stresses. Aggregation was measured using size-exclusion chromatography and PTM levels were calculated using peptide mapping. By combining principal component analysis (PCA) with Raman spectroscopy and circular dichroism (CD) spectroscopy structural analysis we were able to separate proteins based on PTMs and degradation. Furthermore, by identifying key bands that lead to the PCA separation we could correlate spectral peaks to specific PTMs. In particular, we have identified a peak which exhibits a shift in samples with higher levels of Trp oxidation. Through separation of IgG4 aggregates, by size, we have shown a linear correlation between peak wavenumbers of specific functional groups and amount of aggregate present. We therefore demonstrate the capability for Raman spectroscopy to be used as an analytical tool to measure degradation and PTMs in-line with therapeutic production.

M3 - Article

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

ER -