Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry

Steve Hessmann; Cyrille Chery; Anne-Sophie Sikora; Annick Gervais; Christine Carapito

doi:10.1016/j.jpha.2023.03.009

Volume 13 Issue 5

May 2023

Turn off MathJax

Article Contents

Article Navigation > Journal of Pharmaceutical Analysis > 2023 > 13(5): 494-502

Steve Hessmann, Cyrille Chery, Anne-Sophie Sikora, Annick Gervais, Christine Carapito. Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry[J]. Journal of Pharmaceutical Analysis, 2023, 13(5): 494-502. doi: 10.1016/j.jpha.2023.03.009

Citation:

Steve Hessmann, Cyrille Chery, Anne-Sophie Sikora, Annick Gervais, Christine Carapito. Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry[J]. Journal of Pharmaceutical Analysis, 2023, 13(5): 494-502. doi: 10.1016/j.jpha.2023.03.009

Citation:

Steve Hessmann, Cyrille Chery, Anne-Sophie Sikora, Annick Gervais, Christine Carapito. Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry[J]. Journal of Pharmaceutical Analysis, 2023, 13(5): 494-502. doi: 10.1016/j.jpha.2023.03.009

PDF( 1289 KB)

Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry

doi: 10.1016/j.jpha.2023.03.009

a. BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC UMR 7178, University of Strasbourg, CNRS, ProFI-FR2048, Strasbourg, 67000, France;
b. Department of Analytical Development Sciences for Biologicals, UCB Pharma S. A., Braine l'Alleud, 1420, Belgium

Funds:

This project was supported by the “Association Nationale de la Recherche et de la Technologie” and UCB Pharma S.A. (Belgium and France) via the CIFRE fellowship of Steve Hessmann. This work was supported by the “Agence Nationale de la Recherche” via the French Proteomic Infrastructure ProFI FR2048 (ANR-10-INBS-08-03). The authors thank Laura Herment and Tanguy Fortin from Anaquant for their support in the use of the HCP Profiler standard.

Received Date: Mar. 14, 2022
Accepted Date: Mar. 25, 2023
Rev Recd Date: Feb. 20, 2023
Publish Date: Mar. 31, 2023

Abstract

Abstract

Monitoring of host cell proteins (HCPs) during the manufacturing of monoclonal antibodies (mAb) has become a critical requirement to provide effective and safe drug products. Enzyme-linked immunosorbent assays are still the gold standard methods for the quantification of protein impurities. However, this technique has several limitations and does, among others, not enable the precise identification of proteins. In this context, mass spectrometry (MS) became an alternative and orthogonal method that delivers qualitative and quantitative information on all identified HCPs. However, in order to be routinely implemented in biopharmaceutical companies, liquid chromatography-MS based methods still need to be standardized to provide highest sensitivity and robust and accurate quantification. Here, we present a promising MS-based analytical workflow coupling the use of an innovative quantification standard, the HCP Profiler solution, with a spectral library-based data-independent acquisition (DIA) method and strict data validation criteria. The performances of the HCP Profiler solution were compared to more conventional standard protein spikes and the DIA approach was benchmarked against a classical data-dependent acquisition on a series of samples produced at various stages of the manufacturing process. While we also explored spectral library-free DIA interpretation, the spectral library-based approach still showed highest accuracy and reproducibility (coefficients of variation < 10%) with a sensitivity down to the sub-ng/mg mAb level. Thus, this workflow is today mature to be used as a robust and straightforward method to support mAb manufacturing process developments and drug products quality control.
- Host cell proteins,
- Absolute quantification standards,
- Data-independent acquisition

FullText(HTML)

References(32)

References

A.L. Grilo, A. Mantalaris, The increasingly human and profitable monoclonal antibody market. Trends Biotechnol. 37 (2019) 9-16.

H. Kaplon, A. Chenoweth, S. Crescioli, et al., Antibodies to watch in 2022. mAbs 14 (2022), 2014296.

M. Suzuki, C. Kato, A. Kato, Therapeutic antibodies: Their mechanisms of action and the pathological findings they induce in toxicity studies, J. Toxicol. Pathol. 28 (2015) 133-139.

T. Cui, B. Chi, J. Heidbrink Thompson, et al., Cathepsin D: Removal strategy on protein A chromatography, near real time monitoring and characterisation during monoclonal antibody production, J. Biotechnol. 305 (2019) 51-60.

B. Yang, W. Li, H. Zhao, et al., Discovery and characterization of CHO host cell protease-induced fragmentation of a recombinant monoclonal antibody during production process development, J. Chromatogr. B 1112 (2019) 1-10.

R. Molden, M. Hu, E.S. Yen, et al., Host cell protein profiling of commercial therapeutic protein drugs as a benchmark for monoclonal antibody-based therapeutic protein development, mAbs 13 (2021), 1955811.

X. Gao, B. Rawal, Y. Wang, et al., Targeted Host Cell Protein Quantification by LC-MRM enables biologics processing and product characterization, Anal. Chem. 92 (2020) 1007-1015.

X. Li, Y. An, J. Liao, et al., Identification and characterization of a residual host cell protein hexosaminidase B associated with N-glycan degradation during the stability study of a therapeutic recombinant monoclonal antibody product, Biotechnol. Prog. 37 (2021), e3128.

S.S. Rane, R.J. Dearman, I. Kimber, et al., Impact of a heat shock protein impurity on the immunogenicity of biotherapeutic monoclonal antibodies, Pharm. Res. 36 (2019), 51.

H. Falkenberg, D.M. Waldera-Lupa, M. Vanderlaan, et al., Mass spectrometric evaluation of upstream and downstream process influences on host cell protein patterns in biopharmaceutical products, Biotechnol. Prog. 35 (2019), e2788.

D.S. Chahar, S. Ravindran, S.S. Pisal, Monoclonal antibody purification and its progression to commercial scale, Biologicals 63 (2020) 1-13.

K.N. Valente, N.E. Levy, K.H. Lee, et al., Applications of proteomic methods for CHO host cell protein characterization in biopharmaceutical manufacturing, Curr. Opin. Biotechnol. 53 (2018) 144-150.

J. Zhu-Shimoni, C. Yu, J. Nishihara, et al., Host cell protein testing by ELISAs and the use of orthogonal methods, Biotechnol. Bioeng. 111 (2014) 2367-2379.

M. Vanderlaan, J. Zhu-Shimoni, S. Lin, et al., Experience with host cell protein impurities in biopharmaceuticals, Biotechnol. Prog. 34 (2018) 828-837.

N. Pythoud, J. Bons, G. Mijola, et al., Optimized sample preparation and data processing of data-independent acquisition methods for the robust quantification of trace-level host cell protein impurities in antibody drug products, J. Proteome Res. 20 (2021) 923-931.

D.E. Walker, F. Yang, J. Carver, et al., A modular and adaptive mass spectrometry-based platform for support of bioprocess development toward optimal host cell protein clearance, mAbs 9 (2017) 654-663.

F. Yang, D.E. Walker, J. Schoenfelder, et al., A 2D LC-MS/MS strategy for reliable detection of 10-ppm level residual host cell proteins in therapeutic antibodies, Anal. Chem. 90 (2018) 13365-13372.

F. Zhang, W. Ge, G. Ruan, et al., Data-independent acquisition mass spectrometry-based proteomics and software tools: A glimpse in 2020, Proteomics 20 (2020), e1900276.

G. Husson, A. Delangle, J. O’Hara, et al., Dual data-independent acquisition approach combining global hcp profiling and absolute quantification of key impurities during bioprocess development, Anal. Chem. 90 (2018) 1241-1247.

J.C. Silva, M.V. Gorenstein, G.-Z. Li, et al., Absolute quantification of proteins by LCMSE: A virtue of parallel MS acquisition, Mol. Cell. Proteomics 5 (2006) 144-156.

C.E. Doneanu, A. Xenopoulos, K. Fadgen, et al., Analysis of host-cell proteins in biotherapeutic proteins by comprehensive online two-dimensional liquid chromatography/mass spectrometry, mAbs 4 (2012) 24-44.

D. Li, A. Farchone, Q. Zhu, et al., Fast, robust, and sensitive identification of residual host cell proteins in recombinant monoclonal antibodies using sodium deoxycholate assisted digestion, Anal. Chem. 92 (2020) 11888-11894.

M. Trauchessec, A.M. Hesse, A. Kraut, et al., An innovative standard for LC-MS-based HCP profiling and accurate quantity assessment: Application to batch consistency in viral vaccine samples, Proteomics 21 (2021), e2000152.

E.W. Deutsch, A. Csordas, Z. Sun, et al., The ProteomeXchange consortium in 2017: Supporting the cultural change in proteomics public data deposition, Nucleic Acids Res. 45 (2017) D1100-D1106.

D. Bouyssie, A.M. Hesse, E. Mouton-Barbosa, et al., Proline: An efficient and user-friendly software suite for large-scale proteomics, Bioinformatics 36 (2020) 3148-3155.

N. Aboulaich, W.K. Chung, J.H. Thompson, et al., A novel approach to monitor clearance of host cell proteins associated with monoclonal antibodies, Biotechnol. Prog. 30 (2014) 1114-1124.

J. Pezzini, G. Joucla, R. Gantier, et al., Antibody capture by mixed-mode chromatography: A comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins, J. Chromatogr. A 1218 (2011) 8197-8208.

B. Tran, V. Grosskopf, X. Wang, et al., Investigating interactions between phospholipase B-Like 2 and antibodies during Protein A chromatography, J. Chromatogr. A 1438 (2016) 31-38.

N.E. Levy, K.N. Valente, L.H. Choe, et al., Identification and characterization of host cell protein product-associated impurities in monoclonal antibody bioprocessing, Biotechnol. Bioeng. 111 (2014) 904-912.

C. Camacho, G. Coulouris, V. Avagyan, et al., BLAST+: architecture and applications, BMC Bioinformatics 10 (2009), 421.

A. Farrell, S. Mittermayr, B. Morrissey, et al., Quantitative host cell protein analysis using two dimensional data independent LC-MSE, Anal. Chem. 87 (2015) 9186-9193.

S. Kreimer, Y. Gao, S. Ray, et al., Host cell protein profiling by targeted and untargeted analysis of data independent acquisition mass spectrometry data with parallel reaction monitoring verification, Anal. Chem. 89 (2017) 5294-5302.

Relative Articles

Supplements(0)

Cited By

Proportional views