Meet Michael Tran
Graduation Year: May 2017
Field of Study: Pathology
Co- and post-translational processing of a protein often leads to hundreds of proteoforms. Top-down—intact protein—mass spectrometry (TDMS) is valuable for characterization of proteoform diversity in simple mixtures. However, conventional omics-style workflows are protein-centric and interrogate mixtures by multidimensional chromatography that has limited proteoform-level observational range (averaging <5 proteoforms per observed protein). We have shown that off-gel isoelectric focusing (IEF), superficially porous liquid chromatography, and Fourier transform MS (IEF-SPLC-MS) is useful for both protein and proteoform-level screens. Here we will examine fundamentals of IEF-SPLC-MS in a scientific workflow designed to maximize the balance between protein and proteoform investigations, specifically exploring the utility of narrow immobilized pH gradients (IPG) to increase observational range for proteoforms in simple and complex mixtures.
Working in the lab was much more careful planning and preparation than I thought it would be. Many things I took for granted became problematic and I had to figure out a way to circumvent these issues. I became more careful about planning experiments and making sure all the neccesary components were ready and available to ensure everything ran smoothly. One of the biggest things I learned is just because you do not have a lot of raw data or that the data doesn't look like you expected doesn't mean you can't make something out of it.
One of the biggest things I learned is just because you do not have a lot of raw data or that the data doesn't look like you expected doesn't mean you can't make something out of it.
Always make sure you write down everything; write down when you performed an experiment, how it was labelled, and anything you think may be important a few months down the line. Nothing is more frustrating than having samples you're not entirely sure what you did to it.