MassIVE MSV000094082

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Development of an Efficient, Effective, and Economical Technology for Proteome Analysis_part II

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Description

Proteomics experiments often have unreasonably high economic and technical barriers to broad biomedical scientists, which not only result in costly supplies, but also the difficulties to standardize and the reluctance to adopt new techniques. We present an efficient and effective, yet economical approach (named E3technology), for proteomics sample preparation. We developed a novel Empore membrane, which could be used as a robust and low-cost medium to generate LCMS-friendly samples in a rapid and reliable fashion. Using different formats of E3technology, we explored a variety of sample types in varied complexity, quantity, volume, and size. We benchmarked their performance against several established approaches, and our data suggest that E3technology provides equivalent or better performance in terms of proteome-wide identification and quantitation. Moreover, we developed an enhanced yet simplified single vessel approach, E4technology, which opens new avenues for low-input or low-cell proteomics analysis. To further demonstrate their practical applicability, we applied the E3 and E4 technologies to investigate RNA-binding proteins derived from affinity purification, and to profile the intact bacterial cell proteome. Overall, our data suggest that these technologies are highly reliable, widely applicable, easily scalable, and much affordable and feasible to non-expert proteomics laboratories. They represent a breakthrough innovation in biomedical science, and we anticipate their widespread adoption by the proteomics community. [doi:10.25345/C5T14V113] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Proteomics ; on-filter digestion ; in-cell digestion ; E3technology ; E4technology

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Principal Investigators:
(in alphabetical order) 		Yanbao Yu, Department of Chemistry and Biochemistry, University of Delaware, USA, USA
Submitting User: yayu
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