New Findings on Chromatography from Institute for Molecular Biology Summarized (A downstream process allowing the efficient isolation of a recombinant amphiphilic protein from tobacco leaves)
By a News Reporter-Staff News Editor at Life Science Weekly -- Investigators discuss new findings in Laboratory Techniques. According to news reporting from Aachen, Germany, by NewsRx journalists, research stated, "The 65-kDa isoform of human glutamic acid decarboxylase (hGAD65) is a major autoantigen in autoimmune diabetes. The heterologous production of hGAD65 for diagnostic and therapeutic applications is hampered by low upstream productivity and the absence of a robust and efficient downstream process for product isolation."
The news correspondents obtained a quote from the research from Institute for Molecular Biology, "A tobacco-based platform has been developed for the production of an enzymatically-inactive form of the protein (hGAD65mut), but standard downstream processing strategies for plant-derived recombinant proteins cannot be used in this case because the product is amphiphilic. We therefore evaluated different extraction buffers and an aqueous micellar two-phase system (AMTPS) to optimize the isolation and purification of hGAD65mut from plants. We identified the extraction conditions offering the greatest selectivity for hGAD65mut over native tobacco proteins using a complex experimental design approach. Under our optimized conditions, the most efficient initial extraction and partial purification strategy achieved an overall hGAD65mut yield of 92.5% with a purification factor of 12.3 and a concentration factor of 23.8. The process also removed a significant quantity of phenols, which are major contaminants present in tobacco tissue."
According to the news reporters, the research concluded: "This is the first report describing the use of AMTPS for the partial purification of an amphiphilic recombinant protein from plant tissues and our findings could also provide a working model for the initial recovery and partial purification of hydrophobic recombinant proteins from transgenic tobacco plants."
For more information on this research see: A downstream process allowing the efficient isolation of a recombinant amphiphilic protein from tobacco leaves. Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2014;960():34-42. Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences can be contacted at: Elsevier Science Bv, PO Box 211, 1000 Ae Amsterdam, Netherlands (see also Laboratory Techniques).
Our news journalists report that additional information may be obtained by contacting E. Gecchele, Fraunhofer Inst Mol Biol & Appl Ecol IME, Aachen, Germany. Additional authors for this research include S. Schillberg, M. Merlin, M. Pezzotti and L. Avesani.
Keywords for this news article include: Aachen, Europe, Germany, Laboratory Techniques
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