Mass Spectrometry Data Center, Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, Maryland, USA.
Biomolecular Labeling Laboratory, Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, Rockville, Maryland, USA.
Biotechnol Bioeng. 2021 Apr;118(4):1491-1510. doi: 10.1002/bit.27661. Epub 2021 Feb 2.
This paper reports the first implementation of a new type of mass spectral library for the analysis of Chinese hamster ovary (CHO) cell metabolites that allows users to quickly identify most compounds in any complex metabolite sample. We also describe an annotation methodology developed to filter out artifacts and low-quality spectra from recurrent unidentified spectra of metabolites. CHO cells are commonly used to produce biological therapeutics. Metabolic profiles of CHO cells and media can be used to monitor process variability and look for markers that discriminate between batches of product. We have created a comprehensive library of both identified and unidentified metabolites derived from CHO cells that can be used in conjunction with tandem mass spectrometry to identify metabolites. In addition, we present a workflow that can be used for assigning confidence to a NIST MS/MS Library search match based on prior probability of general utility. The goal of our work is to annotate and identify (when possible), all liquid chromatography-mass spectrometry generated metabolite ions as well as create automatable library building and identification pipelines for use by others in the field.
本文报道了一种新型的用于分析中国仓鼠卵巢（CHO）细胞代谢物的质谱文库的首次实现，该文库使用户能够快速鉴定任何复杂代谢物样品中的大多数化合物。我们还描述了一种注释方法，该方法用于从代谢物反复出现的未识别光谱中过滤出伪影和低质量光谱。CHO 细胞常用于生产生物疗法药物。CHO 细胞和培养基的代谢谱可用于监测过程变异性，并寻找可区分产品批次的标志物。我们已经创建了一个综合的 CHO 细胞衍生的已识别和未识别代谢物文库，可与串联质谱联用以鉴定代谢物。此外，我们还提出了一种工作流程，可用于根据一般用途的先验概率为 NIST MS/MS 库搜索匹配分配置信度。我们工作的目标是注释和识别（在可能的情况下）所有液相色谱-质谱生成的代谢物离子，并创建可自动化的文库构建和鉴定管道，以供该领域的其他人使用。
