Development and validation of a UPLC-MS/MS method to quantitate anti-PD1 monoclonal antibody (Toripalimab), and comparison with electrochemiluminescence immunoassay.

J Pharm Biomed Anal

Clinical Pharmacology Research Center, Peking Union Medical College Hospital, State Key Laboratory of Complex Severe and Rare Diseases, NMPA Key Laboratory for Clinical Research and Evaluation of Drug, Beijing Key Laboratory of Clinical PK & PD Investigation for Innovative Drugs, Chinese Academy of Medical Sciences & Peking Union Medical College, No.41, Damucang Hutong, Xicheng District, Beijing 100032, China. Electronic address:

Published: February 2022

AI Article Synopsis

  • Toripalimab is a monoclonal antibody targeting programmed death receptor-1, showing promise in treating various cancers, but there's currently limited methodology for its quantification, mainly relying on electrochemiluminescence immunoassay (ECLIA).
  • Researchers developed a sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method, which involved a detailed five-step sample preparation process to accurately measure toripalimab levels in plasma.
  • The study, which analyzed 77 samples from patients in a phase I trial, found that UPLC-MS/MS reported significantly higher toripalimab levels compared to ECLIA, indicating that UPLC-MS/MS could be a valuable tool for quant

Article Abstract

Toripalimab, a humanized IgG4 monoclonal antibody (mAb) against programmed death receptor-1, is being extensively studied to treat various malignancies. At present, there is no complete methodology reported for quantifying toripalimab, except for an electrochemiluminescence immunoassay (ECLIA) mentioned in several clinical studies. Therefore, a sensitive and robust ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed to accurately detect toripalimab levels, compared with the ECLIA. Plasma samples were pretreated by a five-step process, encompassing denaturation, reduction, alkylation, enzymatic hydrolysis and quenching. And a unique, sensitive and stable enzymatic peptide (ASGYTFTDYEMHWVR) selected as surrogate of toripalimab was eluted and monitored by UPLC-MS/MS system with the linear range of 5.0375-201.5 μg/mL. After fully validated, the UPLC-MS/MS method was applied to determine 77 plasma samples from 29 patients in a phase I clinical trial, and compared with ECLIA based on 56 samples. Wilcoxon paired samples test showed toripalimab levels by UPLC-MS/MS were significantly higher than that by ECLIA (p < 0.001), though a strong correlation was observed (r = 0.96). Moreover, Passing-Bablok regression analysis exhibited constant and proportional biases: UPLC-MS/MS = 2.25 + 1.21 * ECLIA. This discrepancy could be mainly attributed to different forms determined: total mAb for UPLC-MS/MS and free mAb for ECLIA, respectively. As a result, this UPLC-MS/MS method may be complementary to ECLIA to monitor different forms of toripalimab. Beyond that, it can be easily modified to simultaneously quantitate multiple-analyte with a small volume of plasma.

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Source
http://dx.doi.org/10.1016/j.jpba.2021.114515DOI Listing

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