A.-A. V. Misterova, V. A. Chicherin, A. S. Gerasimov
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Optimization of a Purification Method for the Recombinant Platelet-Derived Growth Factor rhPDGF-BB Expressed in the Methylotrophic Yeast Pichia Pastoris
Recombinant human platelet-derived growth factor rhPDGF-BB is one of the major cytokines that has been approved for medical use. Medical drug becaplermin, which contains rhPDGF-BB, has been approved for neuropathic ulcer and severe skin burns treatment, as well as in periodontal surgery (in combination with osteoconductive matrices). In this article, we sought to optimize the purification process to obtain high purity rhPDGF-BB using methylotrophic yeast Pichia pastoris, the production host for rhPDGF-BB. A faster and simpler chromatography purification method has been suggested that allows one to obtain rhPDGF-BB with a purity of more than 98% as determined by SDS-PAGE containing host cell proteins (HCPs) at 33 ± 4 ng/mg, as measured by ELISA. The effective proliferative dose of rhPDGF-BB measured by the WST-1 proliferative assay on 3T3 mouse fibroblast cell culture is 5.02 ± 2.64 ng/mL, which is comparable to commercially available analogues. The optimized method can be attractive for production scale use.
期刊介绍：
Applied Biochemistry and Microbiology is an international peer reviewed journal that publishes original articles on biochemistry and microbiology that have or may have practical applications. The studies include: enzymes and mechanisms of enzymatic reactions, biosynthesis of low and high molecular physiologically active compounds; the studies of their structure and properties; biogenesis and pathways of their regulation; metabolism of producers of biologically active compounds, biocatalysis in organic synthesis, applied genetics of microorganisms, applied enzymology; protein and metabolic engineering, biochemical bases of phytoimmunity, applied aspects of biochemical and immunochemical analysis; biodegradation of xenobiotics; biosensors; biomedical research (without clinical studies). Along with experimental works, the journal publishes descriptions of novel research techniques and reviews on selected topics.
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