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

A.-A. V. Misterova; Мистерова А.-А. В.; V. A. Chicherin; Чичерин В. А.; A. S. Gerasimov; Герасимов А. С.

doi:10.31857/S0555109923040098

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

Authors: Misterova A.V.¹, Chicherin V.A.¹, Gerasimov A.S.¹
Affiliations:
1. Vyatka State University
Issue: Vol 59, No 4 (2023)
Pages: 383-391
Section: Articles
URL: https://ogarev-online.ru/0555-1099/article/view/138794
DOI: https://doi.org/10.31857/S0555109923040098
EDN: https://elibrary.ru/QZLWNT
ID: 138794

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Abstract
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Abstract

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.

Keywords

Human platelet-derived growth factor, growth factors, Pichia pastoris, diabetic foot ulcers, recombinant proteins

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