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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Poly LC PHEA色谱柱,利用p H为7.0的200 mmol·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 complete protein molecular sieve chromatograph tandem mass spectrometry (HPLC-MS / MS) method for the determination of drug conjugate ratio (DAR) based on a cysteine-coupled antibody-conjugated drug (ADC). Methods: The desalted samples were desalted on a Poly LC PHEA column using 200 mmol·L-1 ammonium acetate solution with a pH 7.0 of 7.0. The ADC components containing different numbers of drugs were determined by tandem mass spectrometry. Results: The measured DAR was 4.16 ± 0.10 with a relative standard deviation of 2.40%, and both showed relative proportions of ADC components with varying numbers of drugs. CONCLUSIONS: Complete molecular mass spectrometry coupled with tandem mass spectrometry can determine the fraction of ADCs containing different numbers of drugs and quantitate each component and can be used for DAR analysis of cysteine-coupled ADCs.