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Title: A comprehensive pipeline for translational top-down proteomics from a single blood draw Open Access Deposited

http://creativecommons.org/licenses/by/4.0/
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Abstract
  • Top-down proteomics (TDP) by mass spectrometry (MS) has become increasingly popular in translational spheres as researchers discern the value of proteoform-resolved measurements. The advent of robust prefractionation strategies for intact proteoforms and the emerging availability of high-resolution benchtop mass spectrometers have catalyzed the emergence of high-throughput TDP for the analysis of biological and clinical samples. Here, we provide a comprehensive protocol for TDP of a sample type often investigated in translational research: human peripheral blood mononuclear cells (PBMC). The workflow described herein comprises sample collection, cell lysis, proteoform prefractionation, instrument setup, liquid chromatography-tandem MS (LC-MS/MS) analysis, and data processing, for a minimum completion time of 48 hours. Importantly, this workflow is compatible with multiple PBMC processing strategies and existing biorepository practices, can be used for both small-scale targeted proteoform studies or large-scale multi-sample discovery studies, and has been successfully applied in quantitative contexts for biomarker discovery.
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  • 06/20/2024
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Date issued
  • 2016-11
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  • Forthcoming;Forthcoming
Description sponsorship
  • Research reported in this publication was supported by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Number P41GM108569. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
To Cite this Work:
A comprehensive pipeline for translational top-down proteomics from a single blood draw [Data set]. Indiana University - DataCORE.

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Toby_Nature_Protocols.zip 2024-08-15 2024-08-15 Open Access
manifest.txt 2024-08-16 2024-08-16 282 Bytes Open Access
TobyREADME.txt 2024-08-16 2024-08-16 2.71 KB Open Access

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