Bioproduct Research and Development, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, United States.
Anal Chem. 2024 Oct 22;96(42):16510-16513. doi: 10.1021/acs.analchem.4c00893. Epub 2024 Oct 8.
Host cell proteins (HCPs) are contaminants of biotherapeutics produced from engineered living systems; they can influence the product's quality, efficacy, and toxicity. Liquid chromatography coupled to mass spectrometry can detect HCPs thereby mitigating their risks. However, highly abundant biotherapeutics hamper the detection of low-level HCPs. Sample preparation termed native digestion has proven effective to preferentially digest and draw out HCPs from intact antibodies. Here, we adapted native digestion to adeno-associated viruses (AAV), which is a vector gaining popularity for gene therapy. We leveraged quantitative proteomics using capillary-flow liquid chromatography-mass spectrometry (LC-MS) and demonstrated that native digestion was more effective than applying denaturing conditions to extract the HCPs associated with different AAV serotypes.
宿主细胞蛋白（HCPs）是工程化生物系统生产的生物疗法的污染物；它们会影响产品的质量、疗效和毒性。液相色谱与质谱联用可以检测 HCPs，从而降低其风险。然而，丰度较高的生物疗法会阻碍对低水平 HCPs 的检测。被称为天然消化的样品制备已被证明可有效优先消化和提取完整抗体中的 HCPs。在这里，我们将天然消化法应用于腺相关病毒（AAV），该病毒作为基因治疗的载体越来越受欢迎。我们利用毛细管流动液相色谱-质谱联用（LC-MS）进行定量蛋白质组学分析，结果表明天然消化法比应用变性条件提取不同 AAV 血清型相关的 HCPs 更有效。
