A workflow for absolute quantitation of large therapeutic proteins in biological samples at intact level using LC-HRMS
A workflow for absolute quantitation of large therapeutic proteins in biological samples at intact level using LC-HRMS
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DOI：
10.4155/bio-2016-0096
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发表时间：
2016-08-01
期刊：
影响因子：
1.8
通讯作者：
Weng, Naidong
中科院分区：
文献类型：
--
作者：
Jian, Wenying;Kang, Lijuan;Weng, Naidong
目的：常用的LC-MS工作流程来定量生物样品中的蛋白质治疗剂是“自下而上”的方法。在这项研究中，目的是建立“自上而下”的方法，用于在完整水平上绝对定量生物样品中类似大小的治疗性抗体或蛋白质。材料和方法：使用重组人单克隆抗体作为模型分子，我们提出了一个工作流程，以测量血浆中的大治疗蛋白质在完整的水平上的基础上去卷积高分辨率MS（HRMS）峰。开发了一种新的MultiQuant（TM）软件功能，以自动解卷积峰和处理数据。结果与结论：该工作流具有良好的性能。这是一项概念验证研究，证明了使用LC-HRMS在完整水平上对大型治疗性蛋白质进行生物分析的可行性。
Aim: The commonly used LC-MS workflow to quantify protein therapeutics in biological samples is 'bottom-up' approach. In this study, the aim is to establish 'top-down' approach for absolute quantitation of therapeutic antibodies or proteins of similar sizes in biological samples at intact level. Materials & methods: Using a recombinant human monoclonal antibody as the model molecule, we present a workflow to measure large therapeutic proteins in plasma at intact level based on deconvoluted high-resolution MS (HRMS) peaks. A novel MultiQuant (TM) software function was developed to automatically deconvolute the peaks and process the data. Results & conclusion: The workflow showed satisfying performance. This is a proof of concept study demonstrating the feasibility of bioanalysis of large therapeutic proteins at intact level using LC-HRMS.