Bioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (A*STAR), Centros, Singapore, 138668, Singapore.
Sci Rep. 2024 May 14;14(1):11018. doi: 10.1038/s41598-024-61522-4.
Antibody-drug conjugates (ADC) payloads are cleavable drugs that act as the warhead to exert an ADC's cytotoxic effects on cancer cells intracellularly. A simple and highly sensitive workflow is developed and validated for the simultaneous quantification of six ADC payloads, namely SN-38, MTX, DXd, MMAE, MMAF and Calicheamicin (CM). The workflow consists of a short and simple sample extraction using a methanol-ethanol mixture, followed by a fast liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. The results showed that well-validated linear response ranges of 0.4-100 nM for SN38, MTX and DXd, 0.04-100 nM for MMAE and MMAF, 0.4-1000 nM for CM were achieved in mouse serum. Recoveries for all six payloads at three different concentrations (low, medium and high) were more than 85%. An ultra-low sample volume of only 5 µL of serum is required due to the high sensitivity of the method. This validated method was successfully applied to a pharmacokinetic study to quantify MMAE in mouse serum samples.
抗体药物偶联物 (ADC) 有效载荷是可裂解的药物，作为弹头，在细胞内对癌细胞发挥 ADC 的细胞毒性作用。开发并验证了一种简单而灵敏的工作流程，用于同时定量六种 ADC 有效载荷，即 SN-38、MTX、DXd、MMAE、MMAF 和 Calicheamicin（CM）。该工作流程包括使用甲醇-乙醇混合物进行简短而简单的样品提取，然后进行快速液相色谱串联质谱 (LC-MS/MS) 分析。结果表明，在小鼠血清中，SN38、MTX 和 DXd 的线性响应范围为 0.4-100 nM，MMAE 和 MMAF 的线性响应范围为 0.04-100 nM，CM 的线性响应范围为 0.4-1000 nM，均得到了很好的验证。六种有效载荷在三个不同浓度（低、中、高）下的回收率均超过 85%。由于方法的灵敏度很高，仅需 5 µL 的超低血清量。该经过验证的方法成功应用于一项药代动力学研究，用于定量测定小鼠血清样品中的 MMAE。
