Automated On-tip Affinity Capture Coupled with Mass Spectrometry to Characterize Intact Antibody-Drug Conjugates from Blood.
作者： Ke Sherry Li , Phillip Y. Chu , Aimee Fourie-O’Donohue , Neha Srikumar , Katherine R. Kozak
DOI: 10.1007/S13361-018-1961-7
关键词: Antibody-drug conjugate 、 Chemistry 、 Conjugate 、 Repeatability 、 Proteomics 、 Elution 、 In vitro 、 Mass spectrometry 、 In vivo 、 Chromatography
摘要: Antibody-drug conjugates (ADCs) present unique challenges for ligand-binding assays primarily due to the dynamic changes of drug-to-antibody ratio (DAR) distribution in vivo and vitro. Here, an automated on-tip affinity capture platform with subsequent mass spectrometry analysis was developed accurately characterize DAR ADCs from biological matrices. A variety elution buffers were tested offer optimal recovery, trastuzumab serving as a surrogate ADCs. High assay repeatability (CV 3%) achieved antibody when captured below maximal binding capacity 7.5 μg. Efficient deglycosylation also demonstrated 1 h followed by capture. Moreover, this tip-based affords higher throughput characterization compared well-characterized bead-based method.
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