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. 

Scroll Day! 14th August, 2012

       It has been about 10 months ago when we first met each other. Ice- breaking session-having us introduce ourselves to each other and to the trainers each time a new one comes.  Most of us didn’t know each other, so it was intimidating to a certain extent, slightly awkward and formal.  We had fewer things to share that time– age, preference, academic background and current position in respective jobs, but it didn’t deter us from mingling with one another. Four months flew by. We had great times during class and outside activities, learning and playing together.

       August 14, 2012 arrived soon after. Everyone looked excited, dressed up and polished because it was a big day for us..Finally we were graduating from the BIDP programme after the months of theory and practical classes. This marked the end of Phase One, where each of us received the competency certification from Talent Corp. The days we spent together in the lecture hall daily will also end after the graduation. No more seeing each other every day, having breakfast together, big groups for lunch times, no more hanging out together as often as we did during the class time. 

        In the ceremony hall, the  preparation was intense…everyone was busy. Microphone.. checked, speech and protocol.. checked, photographer..checked, VIP list ... checked , certificate..checked, all the necessities were prepared. Thanks to these people involved (from Talent Corp, Agilent, HAT, I-pharm reps, and invited emcee of the day –Mr. Terrance Dass) the ceremony went swiftly. The ceremony was officiated by none other than the Minister from the Prime Minister’s  Department, Y.Bhg.Tan Sri Nor Mohamed Yakcob, attended by Y.Bhg. Dato Gooi Soon Chai (Agilent), Y.Bhg. Prof Datin Paduka Dr. Khatijah Bt. Mohd Yusoff(MOSTI), Mr Johan Mahmood Merican(Talent Corp), Dato’ Dr. Mohamed Isa Abd Majid  (I-pharm),Mr Azhar Amin (HAT) and invited guests. 

       Eagerly waiting for the arrival of all the VIPs, we saw many photographers and press people coming into the hall one by one..we were all trying to look our best just in case the camera lens might catch us. Suddenly all the photographers and reporters swarmed the building entrance. They’ve arrived!! Giving wide smiles while shaking hands, Tan Sri Nor Mohamed Yakcob was very friendly.  Beside him were Mr. Johan and Dato’ Mohamed Isa, followed by Mr. Azhar . Just moments before that, we had witnessed the arrival of Dato’ Gooi Soon Chai and Prof. Datin Paduka Dr. Khatijah. To be given our certificates in such a big event, we indeed felt special and honoured.

In his speech, Tan Sri mentioned that this program will be a help to achieve what Malaysia aspires in the near future, that is to be a high income nation, as it will provide a high impact talent workforce from local and international university graduates. 


Tan Sri signing the guest book

Emcee for the day, Mr Terrence from Hitz.fm

               Tan Sri Nor speech at Agilent BIDP certificate presentation from Azhar2397

The various candidates from the different organizations represented for in the BIDP
After receiving our certificates and getting pictures taken, Tan Sri spent a few moments talking to us. We found Tan Sri’s spontaneous and humorous side as he joked, and made all of us laugh together with him.  Without any delay, Tan Sri and all the VIPs went to the  Agilent-BIDP Laboratory. He was very impressed to see the lab setting and the equipment inside.

Back in the hall, we congratulated each other, proudly holding our certificates. The next day, we saw the article of us in Bernama news. The experience that we had that day would surely be with us for a long time!


                                                                                       Written by: Nurul Akma & Melanie Ann

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Thought from New Zealand




Chor Ee (4th from left)with BIDP participants

 
The great thing about reflection, especially when trying to remember an event from 6 months ago, is that the memories that come flooding back are the ones that are tied to the strongest emotions. I was asked at the end of 2012 to give feedback on my time with the BIDP students in June covering the topic of DNA detection technologies, more specifically QPCR.

 Leading up to the training itself, I remember being really nervous, maybe with a little stress and anxiety thrown in for good measure. I was coming to train a group of people I had never met, on a brand of instruments that I had never used and in a laboratory I had never seen. Thankfully, Melanie and Nurul from HAT were willing to give up their Saturday to walk me through the lab and the instrumentation, giving me a little more confidence in what I was doing. So, as I reflect on my time with the BIDP, the first mention I would like to make is to appreciate the amazing, professional and dedicated staff at HAT who organised and oversaw the BIDP training programme and who always went the extra mile to make my job as the trainer an easy one.

Chocolates, who does not love chocolates? Figuring that most people like chocolates, I brought some with me from New Zealand to use as an incentive for class participation. However, on the first day of lectures, I was in for a surprise. Not even the tastiest, creamiest chocolate was enough to entice the students to participate in class voluntarily. Thankfully they all had their names on cards in front of them, so encouraging a little class participation was not too hard. Maybe it was the topic being covered, or that a few of the more brave ones had to get the ball rolling, but soon I was being impressed by the calibre of the students, their friendliness, and even their sense of humour, that was present in the lecture room. On a personal note, hearing each team present a QPCR platform and application as a group activity, gave me a sense of pride in them. Their intelligence, ability to communicate effectively and team work was great to watch. I believe they will go far in the careers and Malaysia will be a better place because of their excellence.

Finally, it would be a miss for me not to mention the kind hospitality that was extended to me while in Penang. Having grown up in Penang, it was a real treat to be taken out for lunch and being well fed. My appreciation goes out to Encik Azhar, Mr Choo, Nurul, Mel, Khairul, Eunice and others from HAT and Agilent. It was also an honour to have visits from Talentcorp officials (Puan Norliza Sahar -Senior Manager and Hj Wan Amizan -Manager), who took a personal interest in the BIDP students and the programme, giving time out of their busy schedules to interact with them and to ask some really great questions. It gives me hope to know that there are leaders like these who have the vision to see our nation succeed and who are willing to invest resources into building our future.

Thank you for taking the time to read these simple thoughts. I look forward to the second phase of the BIDP training in February 2013. Drop by for a visit if you are interested, but ask the HAT staff first..."

 
With Mel (Class monitor)
 

By: Chor Ee
(Instructor for qPCR and DNA Detection technology)
 
Chor Ee is a molecular scientist in Wellington Hospital Laboratory Services, New Zealand and a registered scientist with the NZ Medical Laboratory Science Board . Experienced in various industries including molecular diagnostics, genetic research, dairy processing, medical laboratory testing and environmental analysis . An expert in real time PCR assay for the detection of pandemic influenza (H1N1) 2009 virus having adapted the CDC protocol for routine diagnostics