A Sensitive and Controlled Data-Independent Acquisition Method for Proteomic Analysis of Cell Therapies
| Autor: | Camille Lombard-Banek, Kerstin I. Pohl, Edward J. Kwee, John T. Elliott, John E. Schiel |
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| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Journal of Proteome Research. 21:1229-1239 |
| ISSN: | 1535-3907 1535-3893 |
| DOI: | 10.1021/acs.jproteome.1c00887 |
| Popis: | Mass spectrometry (MS)-based proteomic measurements are uniquely poised to impact the development of cell and gene therapies. With the adoption of rigorous instrumental performance qualifications (PQs), large-scale proteomics can move from a research to a manufacturing control tool. Especially suited, data-independent acquisition (DIA) approaches have distinctive qualities to extend multiattribute method (MAM) principles to characterize the proteome of cell therapies. Here, we describe the development of a DIA method for the sensitive identification and quantification of proteins on a Q-TOF instrument. Using the improved acquisition parameters, we defined a control strategy and highlighted some metrics to improve the reproducibility of SWATH acquisition-based proteomic measurements. Finally, we applied the method to analyze the proteome of Jurkat cells that here serves as a model for human T-cells. Raw and processed data were deposited in PRIDE (PXD029780). |
| Databáze: | OpenAIRE |
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