Establishing and Streamlining Workflows for High-pH Reversed-Phase HPLC Fractionation For Proteomics Profiling

Presenter's Name(s)

Lucas Leon

Conference Year

2024

Abstract

High pH reversed phase HPLC fractionation, when coupled to mass spectrometry, is a useful method for covering nearly the entire proteome. This project aimed at establishing such a workflow for the UVM proteomics core to use for future collaborations with universities across the country. A previously homogenized and digested mouse brain was fractionated with traditional methods, and ran on the mass spectrometer. Proteomics based software was then used to determine mouse brain proteome coverage, and total protein count. This was then compared back to samples run via high pH reversed phase HPLC which should display greater proteome coverage.

Primary Faculty Mentor Name

Ying Wai Lam

Graduate Student Mentors

Sydney Cohn-Guthrie

Status

Undergraduate

Student College

College of Arts and Sciences

Program/Major

Biology

Primary Research Category

Life Sciences

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Establishing and Streamlining Workflows for High-pH Reversed-Phase HPLC Fractionation For Proteomics Profiling

High pH reversed phase HPLC fractionation, when coupled to mass spectrometry, is a useful method for covering nearly the entire proteome. This project aimed at establishing such a workflow for the UVM proteomics core to use for future collaborations with universities across the country. A previously homogenized and digested mouse brain was fractionated with traditional methods, and ran on the mass spectrometer. Proteomics based software was then used to determine mouse brain proteome coverage, and total protein count. This was then compared back to samples run via high pH reversed phase HPLC which should display greater proteome coverage.