A highly sensitive and robust LC-MS platform for host cell protein characterization in biotherapeutics

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2023-05-01 DOI:10.1016/j.biologicals.2023.101675

Qinqin Ji, Izabela Sokolowska, Rui Cao, Yulei Jiang, Jingjie Mo, Ping Hu

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引用次数: 0

Abstract

Host cell proteins (HCPs) are a major class of process-related impurities that need to be closely monitored during the production of biotherapeutics. Mass spectrometry (MS) has emerged as a promising tool for HCP analysis due to its specificity for individual HCP's identification and quantitation. However, utilization of MS as a routine characterization tool is still limited due to the time-consuming procedures, non-standardized instrumentation and methodologies, and the limited sensitivity compared to the enzyme-linked immunosorbent assays (ELISA). In this study, we introduced a sensitive (limit of detection (LOD) at 1–2 ppm) and robust HCP profiling platform method with suitable precision and accuracy that can be readily adopted to antibodies and other biotherapeutic modalities without the need for HCP enrichment. The NIST mAb and multiple in-house antibodies were analyzed, and results were benchmarked with other reported studies. In addition, a targeted analysis method with optimized sample preparation for absolute quantitation of lipases was developed and qualified with an LOD of 0.6 ppm and precision of <15%, which can be further improved to an LOD of 5 ppb by using the nano-flow LC.

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一种用于生物治疗中宿主细胞蛋白质表征的高灵敏度和强大的LC-MS平台

宿主细胞蛋白（HCP）是一类主要的工艺相关杂质，在生物治疗药物的生产过程中需要密切监测。质谱（MS）由于其对单个HCP的鉴定和定量的特异性，已成为HCP分析的一种有前途的工具。然而，由于耗时的程序、非标准化的仪器和方法，以及与酶联免疫吸附测定（ELISA）相比灵敏度有限，MS作为常规表征工具的利用仍然有限。在这项研究中，我们引入了一种灵敏（检测限（LOD）为1–2 ppm）和稳健的HCP分析平台方法，该方法具有适当的精度和准确性，可以很容易地用于抗体和其他生物治疗模式，而无需富集HCP。对NIST单克隆抗体和多种内部抗体进行了分析，并将结果与其他报道的研究进行了对比。此外，开发了一种具有用于脂肪酶绝对定量的优化样品制备的靶向分析方法，并以0.6ppm的LOD和<；15%，可以通过使用纳米流LC将其进一步提高到5ppb的LOD。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.