单克隆抗体
计算生物学
质谱法
生物分析
糖基化
化学
抗体
抗体-药物偶联物
色谱法
计算机科学
生物
生物化学
遗传学
作者
Sara Rosati,Yang Yang,Arjan Barendregt,Albert J. R. Heck
出处
期刊:Nature Protocols
[Springer Nature]
日期:2014-03-27
卷期号:9 (4): 967-976
被引量:110
标识
DOI:10.1038/nprot.2014.057
摘要
The molecular complexity of biopharmaceuticals puts severe demands on the bioanalytical techniques required for their comprehensive structural characterization. Mass spectrometry (MS) has gained importance in the analysis of biopharmaceuticals, taking different complementary approaches ranging from peptide-based sequencing to direct analysis of intact proteins and protein assemblies. In this protocol, we describe procedures optimized to perform the analysis of monoclonal antibodies (mAbs) at the intact protein level under pseudo-native conditions, using native MS. Some of the strengths of native MS in the analysis of biopharmaceuticals are its analysis speed, sensitivity and specificity: for most experiments, the whole protocol requires one working day, whereby tens of samples can be analyzed in a multiplexed manner, making it suitable for high-throughput analysis. This method can be used for different applications such as the analysis of mixtures of mAbs, drug-antibody conjugates and the analysis of mAb post-translational modifications, including the qualitative and quantitative analysis of mAb glycosylation.
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