完整蛋白分子筛色谱串联质谱测定药物抗体偶联比

目的:建立完整蛋白分子筛色谱串联质谱分析方法对一种基于半胱氨酸偶联的抗体偶联药物(ADC)的药物抗体偶联比(DAR)进行测定。方法:采用PolyLC PHEA色谱柱,利用pH为7.0的200mmol·L^-1乙酸铵溶液为流动相对脱糖样品进行脱盐,通过串联质谱对含有不同数目药物的ADC组分进行测定。结果:所测定的DAR为4.16±0.10,相对标准差为2.40%,且均能显示含不同数目药物的ADC组分的相对比例。结论:完整蛋白分子筛色谱串联质谱分析可确定ADC中含不同数目药物的组分并对各组分进行定量,可用于基于半胱氨酸偶联的ADC的DAR分析。

Objective:To establish a method of intact size-exclusion chromatography coupled with mass spectrometry for the determination of drug antibody ratio(DAR)of an antibody drug conjugate(ADC)based on cysteine conjugation. Methods:200 mmol·L^-1 ammonium acetate solution(pH 7.0)was used as the mobile phase, and desalting was performed with PHEA chromatographic column of PolyLC Inc. The mass spectrometric analysis was performed to determine the DAR of the ADC supsets. Results:The DAR was measured as 4.16±0.10 with the relative standard deviation of 2.40%. Furthermore, the relative ratio of ADC supsets containing different number of drugs could also be obtained. Conclusion:The analytical method of intact size-exclusion chromatography coupled with mass spectrometry in the study can be applied to the quantitative analysis of ADC supsets containing different numbers of drugs, and the method can be successfully applied to the determination of the DAR of ADC based on cysteine conjugation.

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