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

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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.

Graphical abstract

Highlights

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Multi-epitope affinity technology showed superior enrichment over mono-epitope.
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A bioanalytical platform was developed utilizing multi-epitope affinity technology.
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In vitro models were established to predict trastuzumab modification trends.
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Modification levels of trastuzumab in patients' sera were monitored in vivo.

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Author and article information

Contributors

Jia-Huan Qu

Zhengjin Jiang

Qiqin Wang

Journal

Journal ID (nlm-ta): J Pharm Anal

Journal ID (iso-abbrev): J Pharm Anal

Title: Journal of Pharmaceutical Analysis

Publisher: Xi'an Jiaotong University

ISSN (Print): 2095-1779

ISSN (Electronic): 2214-0883

Publication date PMC-release: 04 June 2024

Publication date (Print): November 2024

Publication date (Electronic): 04 June 2024

Volume: 14

Issue: 11

Electronic Location Identifier: 101015

Affiliations

[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, Guang dong, 518107, China

[e ]Guangdong Institute for Drug Control, Guangzhou, 510663, China

Author notes

[* ]Corresponding author. qiqinxtu@ 123456163.com

[** ]Corresponding author. jzjjackson@ 123456hotmail.com

[*** ]Corresponding author. jiahuanqu@ 123456jnu.edu.cn

[1]

Both authors contributed equally to this work.

Article

Publisher Item ID: S2095-1779(24)00112-6 Publisher ID: 101015

DOI: 10.1016/j.jpha.2024.101015

PMC ID: 11652880

PubMed ID: 39698314

SO-VID: 9a50c85a-3ea7-4c9b-94c9-f27083f259d0

License:

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

History

Date received : 14 March 2024

Date revision received : 19 May 2024

Date accepted : 30 May 2024

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