Purification Method Optimization of Recombinant Platelet-Derived Growth Factor rhPDGF-BB Expressed in Methylotrophic Yeast Pichia pastoris

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Resumo

Recombinant human platelet-derived growth factor rhPDGF-BB is one of the major cytokines, which has been approved for medical use. Medical drug “becaplermin”, containing 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 purification process to obtain high purity rhPDGF-BB using methylotrophic yeast Pichia pastoris – a production host for rhPDGF-BB. A faster and simpler chromatography purification method has been suggested which allows to obtain rhPDGF-BB with purity >98% as determined by SDS-PAGE and containing host cell proteins (HCP) 33 ± 4 ng/mg, as measured by ELISA. The effective proliferative dose of rhPDGF-BB measured by 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.

Sobre autores

A.-A. Misterova

Vyatka State University

Autor responsável pela correspondência
Email: usr21438@vyatsu.ru
Russia, 610000 , Kirov

V. Chicherin

Vyatka State University

Email: usr21438@vyatsu.ru
Russia, 610000 , Kirov

A. Gerasimov

Vyatka State University

Email: usr21438@vyatsu.ru
Russia, 610000 , Kirov

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Declaração de direitos autorais © А.-А.В. Мистерова, В.А. Чичерин, А.С. Герасимов, 2023