Understanding LC-MS in Proteomics

       This set of classes lasted from the 6th to the 14th of Dec 2012. It had been sometime since we had any lab practical sessions, this was the first and last in Phase II of the BIDP programme. Prof Manfred, who has a very wide knowledge of LC-MS in proteomics, came all the way from Germany and settled in Singapore.  We anticipated to learn much from such an experienced person whose has had hands-on work for more than 20 years.

   In the first two days, we learned the very fundamentals in proteomics and chromatography in research. Our memories were refreshed on the knowledge of HPLC as we had sat through trainings prior to that. While we were taught the technical specs of chromatography and HPLC previously, Prof covered in-depth about HPLC methods in proteomics research.

Prof Manfred in class

          Proteomics, being a study about the protein structure and function, Prof taught us of the chronological evolution of protein research with the approach used to study protein. Proteins are big and complex molecules, analysed to get the sequence, the folding, the function, the binding and even the inhibition of its function. We received some insights of the work in the field of protein research as Prof generously shared his experiences of handling and troubleshooting experiments.

       The next 5 days of training were spent in practical training. Prof had brought along some standards, samples and consumables to be used.  After groups were formed, we had the privilege to bring in our samples of choice to be tested. Prof said he would like to use samples that he had not tested before and wanted to buy local fruits to be sampled! It was interesting.. We tested mangosteen, Roselle, Hibiscus and even the crude sea cucumber extract which Wan Ting had brought in for us. Thank you Wan Ting!! That was indeed rare and special.

Prof Manfred demonstrating to us in the lab

          During the practical times, we were joined by Ivan as our tutor. He helped us maintain and troubleshoot the UHPLC machine and even set up the protocol on the machine for us. Running these kinds of samples on the UHPLC on was not as fast as simple pharmaceutical products which were ‘cleaner’.  It took roughly 20 minutes per run for each vial due to gradient concentration of the buffer that was needed to separate the compound… 

Protein sample is complex remember? And it was even more complex using the crude sample that we had. We had not selectively choose any protein to elute out. The result??? Many peaks were obtained, but not identified, as that would be a whole training by itself. But the interesting part was of course the sample prep time (hands-on time!). Grinding, blending, pipetting, filtering (where the clean lab become messy. Again) Prof was very kind in showing us how to prepare the samples and even prepared the buffer himself.

          Even with the practical sessions going on well, we still had our theory sessions in class (in the morning before we went to the lab). Prof taught us on Mass Spectrometry (MS), the principle, the parts, the result analysis and the requirement to run protein samples. Generally, we learnt how to run an assay in MS. Prof shared a few journals for us to look into and the reference samples to use in protein research. There’s really so much to explore. Proteins are only a small portion of research in understanding our cell-networking, or what we call biological system. That’s what makes the proteomic research on-going and expanding. 

Thank you Prof, we have learned much!  ^.^

                                                                                                                        Written by: Nurul Akma

Prof. Manfred Raida
PhD in Biochemistry, on the chemical modification of the anion transport protein of the red blood cell membrane in Johann Wolfgang Goethe-Universität Frankfurt am Main
The founder of Proteomics Asia Pte. Ltd., involved in the Society for Mass Spectrometry (Singapore) and the National University of Singapore (NUS), Prof Manfred is a specialist in developing and deploying new methods in the protein identification, targeted proteomics and chemical proteomics. He itakes special interest in protein identification methods using mass spectrometry as well as separation science.