PDF(Pubmed)
Abstract：
Antibodies are one of the most formidable molecular weapons available to our immune system. Their high specificity against a target (antigen) and capability of triggering different immune responses (e.g., complement system activation and antibody-dependent cell-mediated cytotoxicity) make them ideal drugs to fight many different human diseases. Currently, both monoclonal antibodies and more complex molecules based on the antibody scaffold are used as biologics. Naturally, such highly heterogeneous molecules require dedicated analytical methodologies for their accurate characterization. Mass spectrometry (MS) can define the presence and relative abundance of multiple features of antibodies, including critical quality attributes. The combination of small and large variations within a single molecule can only be determined by analyzing intact antibodies or their large (25 to 100 kDa) subunits. Hence, top-down (TD) and middle-down (MD) MS approaches have gained popularity over the last decade. In this Young Scientist Feature we discuss the evolution of TD and MD MS analysis of antibodies, including the new frontiers that go beyond biopharma applications. We will show how this field is now moving from the \"quality control\" analysis of a known, single antibody to the high-throughput investigation of complex antibody repertoires isolated from clinical samples, where the ultimate goal is represented by the complete gas-phase sequencing of antibody molecules without the need of any a priori knowledge.
摘要：
抗体是我们免疫系统最强大的分子武器之一。它们对靶标（抗原）的高特异性和触发不同免疫反应的能力（例如，补体系统激活和抗体依赖性细胞介导的细胞毒性）使它们成为对抗许多不同人类疾病的理想药物。目前,单克隆抗体和基于抗体支架的更复杂的分子都被用作生物制剂。自然,这种高度异质的分子需要专门的分析方法来准确表征。质谱（MS）可以定义抗体的多个特征的存在和相对丰度，包括关键质量属性。单个分子内的小变异和大变异的组合只能通过分析完整的抗体或它们的大(25至100kDa)亚基来确定。因此，在过去的十年中，自上而下（TD）和中下（MD）MS方法已经普及。在这个年轻的科学家特征中，我们讨论了抗体的TD和MDMS分析的演变，包括超越生物制药应用的新领域。我们将展示这个领域现在是如何从“质量控制”分析已知的，从临床样品中分离的复杂抗体库的高通量研究的单一抗体，其中最终目标是通过抗体分子的完整气相测序来表示，而无需任何先验知识。
