A DAR sensitive hybrid LBA-LC-MS/MS assay for quantitation of conjugated payload from a non-cleavable ADC in human serum.

IF 2.1 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Bioanalysis Pub Date : 2026-08-01 Epub Date: 2026-08-17 DOI:10.1080/17576180.2026.2716471

Yongjun Xue, Eric Ma, Brian Melo, Madhan Masilamani, Yiming Cheng, Jim X Shen, M Shane Woolf, Korrey D Allen, Jordan M Honrine, Michael P Waldron, William R Mylott

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引用次数: 0

Abstract

Background: Quantitating antibody drug conjugates (ADCs) with traditional ligand binding assays (LBAs) requires cytotoxic payload targeting reagents, which may be needed for ADC capture/detection. LBAs are drug antibody ratio insensitive (DAR insensitive) and prone to under- or over-estimating DAR species.

Methods: This hybrid LBA LC MS/MS assay measures concentrations for a novel ADC drug (BMS-X) with an engineered, site‑specific, non-cleavable linker, and a unique payload. Recombinant target protein-extracellular domain fused with mouse IgG-Fc was used for ADC pull down in human serum. Pepsin digestion released unique toxin linker peptide complexes for two conjugation sites.

Results and conclusions: This novel, validated DAR-sensitive hybrid-LCMS PK assay measured payload-conjugated BMS-X levels in human serum, quantitated surrogate analytes for both unique conjugation sites, and was used to support a first-in-human clinical trial.

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一种DAR敏感的混合LBA-LC-MS/MS方法用于定量人血清中不可切割ADC的共轭有效载荷。

背景：用传统的配体结合测定法（LBAs）定量抗体药物偶联物（ADC）需要细胞毒性有效载荷靶向试剂，这可能需要用于ADC的捕获/检测。LBAs对药物抗体比例不敏感（DAR不敏感），容易低估或高估DAR种类。方法：这种混合式LBA LC MS/MS法测量一种新型ADC药物（BMS-X）的浓度，该药物具有工程化的、位点特异性的、不可切割的连接体和独特的有效载荷。利用重组靶蛋白胞外结构域与小鼠IgG-Fc融合的方法，对人血清进行ADC的拉降。胃蛋白酶消化释放出独特的毒素连接肽复合物，用于两个偶联位点。结果和结论：这种新型的、经过验证的dar敏感的混合lcms PK测定方法测量了人血清中有效载荷偶联的BMS-X水平，定量了两种独特偶联位点的替代分析物，并用于支持首次人体临床试验。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.