The TGF‐β cytokine myostatin is considered to be one of the key regulators of skeletal muscle mass. Consequently, specific inhibitors of the growth factor and its signaling pathways are promising therapeutics for the treatment of muscle wasting disorders as well as potential performance‐enhancing agents in sports. Domagrozumab is a humanized monoclonal antibody that neutralizes the circulating cytokine, thus preventing receptor activation. Within this study, two complementary detection assays for Domagrozumab from serum were developed by using ammonium sulfate precipitation and immunoaffinity purification either in combination with tryptic digestion and LC‐HRMS or Western blotting. While the LC‐HRMS assay is highly specific for diagnostic peptides originating from both the heavy and the light chain of the antibody, the second assay is capable of generically detecting intact therapeutic proteins comprising a human Fc domain and exhibiting high specificity for dimeric myostatin/GDF‐11. Following optimization, both assays were comprehensively characterized. They can readily be modified to include further protein drugs and will expand the range of available tests for emerging myostatin inhibitors.

中文翻译：

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通过蛋白质印迹和LC-HRMS检测血清中的肌生长抑制素中和抗体Domagrozumab。

TGF-β细胞因子肌生长抑制素被认为是骨骼肌质量的关键调节剂之一。因此，生长因子及其信号通路的特异性抑制剂有望成为治疗肌肉萎缩症以及运动中潜在性能增强剂的理想疗法。Domagrozumab是一种人源化单克隆抗体，可中和循环中的细胞因子，从而防止受体活化。在这项研究中，通过使用硫酸铵沉淀和免疫亲和纯化结合胰蛋白酶消化和LC-HRMS或Western印迹，开发了两种针对血清中Domagrozumab的互补检测方法。尽管LC‐HRMS分析对于源自抗体重链和轻链的诊断肽具有高度特异性，第二种检测方法能够一般性地检测包含人Fc结构域的完整治疗性蛋白质，并且对二聚体肌肉生长抑制素/ GDF-11表现出高特异性。优化后，对两种测定法进行了全面表征。可以很容易地对其进行修饰，使其包含其他蛋白质药物，并将扩大新兴肌生长抑制素抑制剂的可用检测范围。