Comparative study of trastuzumab modification analysis using mono/multi-epitope affinity technology with LC-QTOF-MS

Chengyi Zuo; Jingwei Zhou; Sumin Bian; Qing Zhang; Yutian Lei; Yuan Shen; Zhiwei Chen; Peijun Ye; Leying Shi; Mao Mu; Jia-Huan Qu; Zhengjin Jiang; Qiqin Wang

doi:10.1016/j.jpha.2024.101015

Volume 14 Issue 11

Nov. 2024

Turn off MathJax

Article Contents

Article Navigation > Journal of Pharmaceutical Analysis > 2024 > 14(11): 101015

Chengyi Zuo, Jingwei Zhou, Sumin Bian, Qing Zhang, Yutian Lei, Yuan Shen, Zhiwei Chen, Peijun Ye, Leying Shi, Mao Mu, Jia-Huan Qu, Zhengjin Jiang, Qiqin Wang. Comparative study of trastuzumab modification analysis using mono/multi-epitope affinity technology with LC-QTOF-MS[J]. Journal of Pharmaceutical Analysis, 2024, 14(11): 101015. doi: 10.1016/j.jpha.2024.101015

Citation:

Chengyi Zuo, Jingwei Zhou, Sumin Bian, Qing Zhang, Yutian Lei, Yuan Shen, Zhiwei Chen, Peijun Ye, Leying Shi, Mao Mu, Jia-Huan Qu, Zhengjin Jiang, Qiqin Wang. Comparative study of trastuzumab modification analysis using mono/multi-epitope affinity technology with LC-QTOF-MS[J]. Journal of Pharmaceutical Analysis, 2024, 14(11): 101015. doi: 10.1016/j.jpha.2024.101015

Citation:

Chengyi Zuo, Jingwei Zhou, Sumin Bian, Qing Zhang, Yutian Lei, Yuan Shen, Zhiwei Chen, Peijun Ye, Leying Shi, Mao Mu, Jia-Huan Qu, Zhengjin Jiang, Qiqin Wang. Comparative study of trastuzumab modification analysis using mono/multi-epitope affinity technology with LC-QTOF-MS[J]. Journal of Pharmaceutical Analysis, 2024, 14(11): 101015. doi: 10.1016/j.jpha.2024.101015

PDF( 3336 KB)

Comparative study of trastuzumab modification analysis using mono/multi-epitope affinity technology with LC-QTOF-MS

doi: 10.1016/j.jpha.2024.101015

a. Institute of Pharmaceutical Analysis, College of Pharmacy/State Key Laboratory of Bioactive Molecules and Druggability Assessment/Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research of China, Jinan University, Guangzhou, 510632, China;
b. School of Engineering, Westlake University, Hangzhou, 310024, China;
c. The First Affiliated Hospital of Jinan University, Guangzhou, 510632, China;
d. School of Biomedical Engineering, Sun Yat-sen University, Shenzhen, Guangdong, 518107, China;
e. Guangdong Institute for Drug Control, Guangzhou, 510663, China

Funds:

This work was supported by the National Natural Science Foundation of China (Grant Nos.: 82373829, 82273893, and 82173773), the Natural Science Foundation of Guangdong Province, China (Grant Nos.: 2021A1515220099, and 2022A1515011576), the High-End Foreign Experts Project, China (Grant No.: G2021199005L), and the Science and Technology Program of Guangdong Provincial Medical Products Administration, China (Grant Nos.: 2023TDZ11, and 2022ZDB04).

Received Date: Mar. 14, 2024
Accepted Date: May 30, 2024
Rev Recd Date: May 19, 2024
Publish Date: Jun. 04, 2024

Abstract

Abstract

Dynamic tracking analysis of monoclonal antibodies (mAbs) biotransformation in vivo is crucial, as certain modifications could inactivate the protein and reduce drug efficacy. However, a particular challenge (i.e. immune recognition deficiencies) in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen. To address this limitation, a multi-epitope affinity technology utilizing the metal organic framework (MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region (CDR) mimotope peptide (HH24) and non-CDR mimotope aptamer (CH1S-6T) onto the surface of MOF@Au nanocomposite. Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology. Moreover, the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry (LC-QTOF-MS). Therefore, multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb. Particularly, the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids. Compared to the spiked phosphate buffer (PB) model, faster modification trends were monitored in the spiked serum and patients' sera due to the catalytic effect of plasma proteins and relevant proteases. Differences in peptide modification levels of trastuzumab in patients' sera were also monitored. In summary, the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb, contributing to improved understanding and paving the way for future research and clinical applications.
- Monoclonal antibody,
- Multi-epitope affinity technology,
- Biotransformation analysis,
- LC-QTOF-MS,
- Breast cancer

FullText(HTML)

References(45)

References

[1]	G. Pantaleo, B. Correia, C. Fenwick, et al., Antibodies to combat viral infections: Development strategies and progress, Nat. Rev. Drug Discov. 21 (2022) 676-696.
[2]	H. Kaplon, S. Crescioli, A. Chenoweth, et al., Antibodies to watch in 2023, mAbs 15 (2023), 2153410.
[3]	D. Corti, L.A. Purcell, G. Snell, et al., Tackling COVID-19 with neutralizing monoclonal antibodies, Cell 184 (2021) 4593-4595.
[4]	P. Zhu, S.-Y. Li, J. Ding, et al., Combination immunotherapy of glioblastoma with dendritic cell cancer vaccines, anti-PD-1 and poly I:C, J. Pharm. Anal. 13 (2023) 616-624.
[5]	J.-J. Zhang, C.-G. Song, M. Wang, et al., Monoclonal antibody targeting mu-opioid receptor attenuates morphine tolerance via enhancing morphine-induced receptor endocytosis, J. Pharm. Anal. 13 (2023) 1135-1152.
[6]	C.T. Walsh, S. Garneau-Tsodikova, G.J. Gatto, Protein posttranslational modifications: The chemistry of proteome diversifications, Angew. Chem. Int. Ed. 44 (2005) 7342-7372.
[7]	J.C. Tran, D. Tran, A. Hilderbrand, et al., Automated affinity capture and on-tip digestion to accurately quantitate in vivo deamidation of therapeutic antibodies, Anal. Chem. 88 (2016) 11521-11526.
[8]	F. Cymer, H. Beck, A. Rohde, et al., Therapeutic monoclonal antibody N-glycosylation-structure, function and therapeutic potential, Biologicals 52 (2018) 1-11.
[9]	Y. Mimura, R. Saldova, Y. Mimura-Kimura, et al., Micro-heterogeneity of antibody molecules, Exp. Suppl. 112 (2021) 1-26.
[10]	C. Nowak, A. Tiwari, H. Liu, Asparagine deamidation in a complementarity determining region of a recombinant monoclonal antibody in complex with antigen, Anal. Chem. 90 (2018) 6998-7003.
[11]	Y. Yan, H. Wei, Y. Fu, et al., Isomerization and oxidation in the complementarity-determining regions of a monoclonal antibody: A study of the modification-structure-function correlations by hydrogen-deuterium exchange mass spectrometry, Anal. Chem. 88 (2016) 2041-2050.
[12]	B. Beyer, M. Schuster, A. Jungbauer, et al., Microheterogeneity of recombinant antibodies: Analytics and functional impact, Biotechnol. J. 13 (2018), 1700476.
[13]	J.A. Pavon, L. Xiao, X. Li, et al., Selective tryptophan oxidation of monoclonal antibodies: Oxidative stress and modeling prediction, Anal. Chem. 91 (2019) 2192-2200.
[14]	A. Beck, E. Wagner-Rousset, D. Ayoub, et al., Characterization of therapeutic antibodies and related products, Anal. Chem. 85 (2013) 715-736.
[15]	N. Yang, Q. Tang, P. Hu, et al., Use of in vitro systems to model in vivo degradation of therapeutic monoclonal antibodies, Anal. Chem. 90 (2018) 7896-7902.
[16]	S. Schadt, S. Hauri, F. Lopes, et al., Are biotransformation studies of therapeutic proteins needed? Scientific considerations and technical challenges, Drug Metab. Dispos. 47 (2019) 1443-1456.
[17]	P. Bults, R. Bischoff, H. Bakker, et al., LC-MS/MS-based monitoring of in vivo protein biotransformation: Quantitative determination of trastuzumab and its deamidation products in human plasma, Anal. Chem. 88 (2016) 1871-1877.
[18]	Y. Li, M. Monine, Y. Huang, et al., Quantitation and pharmacokinetic modeling of therapeutic antibody quality attributes in human studies, mAbs 8 (2016) 1079-1087.
[19]	Y. Lei, Y. Shen, C. Zuo, et al., Emerging affinity ligands and support materials for the enrichment of monoclonal antibodies, Trends Anal. Chem. 157 (2022), 116744.
[20]	M.J. Suh, J.B. Powers, C.M. Daniels, et al., Enhanced pharmacokinetic bioanalysis of antibody-drug conjugates using hybrid immunoaffinity capture and microflow LC-MS/MS, AAPS J. 25 (2023), 68.
[21]	L. Dong, N. Bebrin, K. Piatkov, et al., An automated multicycle immunoaffinity enrichment approach developed for sensitive mouse IgG1 antibody drug analysis in mouse plasma using LC/MS/MS, Anal. Chem. 93 (2021) 6348-6354.
[22]	F. Yin, D. Adhikari, M. Sun, et al., Bioanalysis of an antibody drug conjugate (ADC) PYX-201 in human plasma using a hybrid immunoaffinity LC-MS/MS approach, J. Chromatogr. B. 1223 (2023), 123715.
[23]	C. Zhu, H. Han, Z. Chen, et al., Tetrapeptide-based mimotope affinity monolith for the enrichment and analysis of anti-HER2 antibody and antibody-drug conjugate, Anal. Chim. Acta 1246 (2023), 340892.
[24]	A. Gonzalez-Quintela, R. Alende, F. Gude, et al., Serum levels of immunoglobulins (IgG, IgA, IgM) in a general adult population and their relationship with alcohol consumption, smoking and common metabolic abnormalities, Clin. Exp. Immunol. 151 (2007) 42-50.
[25]	M.A. French, G. Harrison, Serum IgG subclass concentrations in healthy adults: A study using monoclonal antisera, Clin. Exp. Immunol. 56 (1984) 473-475.
[26]	L. Lu, X. Liu, C. Zuo, et al., In vitro/in vivo degradation analysis of trastuzumab by combining specific capture on HER2 mimotope peptide modified material and LC-QTOF-MS, Anal. Chim. Acta 1225 (2022), 340199.
[27]	S. Yin, C.V. Pastuskovas, L.A. Khawli, et al., Characterization of therapeutic monoclonal antibodies reveals differences between in vitro and in vivo time-course studies, Pharm. Res. 30 (2013) 167-178.
[28]	Y. Zhang, S.-L. Wu, Y. Li, Comparative study of profiling post-translational modifications of a circulating antibody drug in human with different capture reagents, Biologicals 45 (2017) 93-95.
[29]	S. Bauer, C. Serre, T. Devic, et al., High-throughput assisted rationalization of the formation of metal organic frameworks in the Iron(III) aminoterephthalate solvothermal system, Inorg. Chem. 47 (2008) 7568-7576.
[30]	Y. Hu, H. Cheng, X. Zhao, et al., Surface-enhanced Raman scattering active gold nanoparticles with enzyme-mimicking activities for measuring glucose and lactate in living tissues, ACS Nano 11 (2017) 5558-5566.
[31]	C. Nowak, A. Tiwari, H. Liu, Asparagine deamidation in a complementarity determining region of a recombinant monoclonal antibody in complex with antigen, Anal. Chem. 90 (2018) 6998-7003.
[32]	R. Zeunik, A.F. Ryuzoji, A. Peariso, et al., Investigation of immune responses to oxidation, deamidation, and isomerization in therapeutic antibodies using preclinical immunogenicity risk assessment assays, J. Pharm. Sci. 111 (2022) 2217-2229.
[33]	Y. Wang, X. Li, Y.-H. Liu, et al., Simultaneous monitoring of oxidation, deamidation, isomerization, and glycosylation of monoclonal antibodies by liquid chromatography-mass spectrometry method with ultrafast tryptic digestion, mAbs 8 (2016) 1477-1486.
[34]	M. Cao, N.D. Mel, J. Wang, et al., Characterization of N-terminal glutamate cyclization in monoclonal antibody and bispecific antibody using charge heterogeneity assays and hydrophobic interaction chromatography, J. Pharm. Sci. 111 (2022) 335-344.
[35]	Z. Liu, Y. Cao, L. Zhang, et al., In-depth characterization of mAb charge variants by on-line multidimensional liquid chromatography-mass spectrometry, Anal. Chem. 95 (2023) 7977-7984.
[36]	Y.D. Liu, A.M. Goetze, R.B. Bass, et al., N-terminal glutamate to pyroglutamate conversion in vivo for human IgG2 antibodies, J. Biol. Chem. 286 (2011) 11211-11217.
[37]	A.B. Joshi, M. Sawai, W.R. Kearney, et al., Studies on the mechanism of aspartic acid cleavage and glutamine deamidation in the acidic degradation of glucagon, J. Pharm. Sci. 94 (2005) 1912-1927.
[38]	Q. He, F. Chen, Z. Zhao, et al., Supramolecular mimotope peptide nanofibers promote antibody-ligand polyvalent and instantaneous recognition for biopharmaceutical analysis, Anal. Chem. 96 (2024) 5940-5950.
[39]	J. Yang, A. Zhou, M. Li, et al., Mimotope peptide modified pompon mum-like magnetic microparticles for precise recognition, capture and biotransformation analysis of rituximab in biological fluids, Acta Pharm. Sin. B 14 (2024) 1317-1328.
[40]	J. Zhou, C. Zuo, H. Tian, et al., Magnetic composite membrane roll column for rapid and high efficiency separation of antibodies, Sep. Purif. Technol. 309 (2023), 123107.
[41]	S. Huang, M. Zhang, F. Chen, et al., A chimeric hairpin DNA aptamer-based biosensor for monitoring the therapeutic drug bevacizumab, Analyst 149 (2024) 212-220.
[42]	Q. Luo, H. Zou, Q. Zhang, et al., High-performance affinity chromatography with immobilization of protein A and L-histidine on molded monolith, Biotechnol. Bioeng. 80 (2002) 481-489.
[43]	W. Liu, A.L. Bennett, W. Ning, et al., Monoclonal antibody capture and analysis using porous membranes containing immobilized peptide mimotopes, Anal. Chem. 90 (2018) 12161-12167.
[44]	L. Huang, J. Lu, V.J. Wroblewski, et al., In vivo deamidation characterization of monoclonal antibody by LC/MS/MS, Anal. Chem. 77 (2005) 1432-1439.
[45]	S. Yin, C.V. Pastuskovas, L.A. Khawli, et al., Characterization of therapeutic monoclonal antibodies reveals differences between in vitro and in vivo time-course studies, Pharm. Res. 30 (2013) 167-178.