Update for Western blot method for protein measurement
(Vienna, 10 Dec. 2010) In medical and biological research the so-called “Western blot” technique is a widely used method for proving the quality and quantity of proteins using corresponding antibodies. A large part of medical research is based on this technique. Univ. Prof. Dr. Gert Lubec from MedUni Vienna is now presenting an extended method which rules out possible inaccuracies in the proof of proteins.
The advantages of the Western blot technique are the rather low costs and the also rather low time required. Probably one of the reasons that this method is widely used for the proof of proteins and also why many studies and research works are based on it. In this process Gert Lubec, head of a renowned protein laboratory at the Department of Paediatrics and Adolescent Medicine at MedUni Vienna, sees the danger that the used antibodies are not specific enough, however, and could also react with other proteins rather than solely with the target protein. This is why, in Lubec’s opinion, the results of the Western blot method must be viewed carefully in the current form and a large part of the work in the past must therefore be called into question. Ultimately this also concerns other tests which work with antibodies, possibly up to therapeutic antibodies.
With this problem in mind Lubec and his team have now developed a procedure which, although also based on a pure immune response (antibody-antigen), reaches a considerably more specific protein chemical level, however. With protein chemical methods it is therefore possible to very precisely identify the target proteins detected by an antibody, even if these occur in modified variations. But this precise method is also associated with extra costs and time, which are also reflected in the costs of better-quality antibodies, of course, and this may prevent the high-grade procedure becoming widespread for a while yet.
Lubec is confident however: “We are working on it and will establish contacts with the companies which produce antibodies. Companies which have reliable specific antibodies will have an advantage on the market, even if it will take time to test thousands of antibodies for their specificity. It will also certainly take time until referees from the scientific journals demand proof of the specificity and not only the company and catalogue number of the used antibody.“
The work presenting the new method for characterising the recognised protein using Western blot and on the 2D gel/mass spectrometry level appeared in the renowned specialist journal “Electrophoresis”:
Generation and characterization of a specific polyclonal antibody against the mouse serotonin receptor 1A: A state-of-the-art recommendation how to characterize antibody specificity
Gert Lubec, Seok Heo
Electrophoresis 2010, DOI 10.1002/elps.201000374
Univ. Prof. Dr. Gert Lubec, born in 1948, studied medicine in Vienna and was already cooperating in biochemical scientific projects during his studies. After graduating Lubec received his own research laboratory at the Paediatric Clinic of MedUni Vienna, he completed his specialist training in paediatrics, and qualified as a professor in a purely protein chemical theme. In recent years he has published studies in the area of neuroproteomics. Gert Lubec has worked as a guest professor in England and France, is a member of the editorial committee of the renowned journal “Proteomics” and editor and founder of the journal “Amino Acids, The forum for Amino Acid and Protein Research” published since 1991 by the Springer publishing house.