Expression and purification of platelet-derived growth factor and evaluation of its function in binding to fibrinogen

Hasannia, Sadegh; Dabirmanesh, Bahareh; Mollasalehi, Maryam

Expression and purification of platelet-derived growth factor and evaluation of its function in binding to fibrinogen

Document Type : Original Research

Authors

Sadegh Hasannia ¹

Bahareh Dabirmanesh ¹

Maryam Mollasalehi ²

¹ Tarbiat Modares university

² Tarbiat Modares university phD Student

Abstract

Abstract

The wound healing process is a complex and dynamic process that involves many metabolic pathways. This process consists of three phases inflammation, cell proliferation, and tissue regeneration stages. Successful wound healing depends on careful regulation and coordination between the factors involved. Until recent years, the strategy of treating chronic wounds was limited to wound preparation, removal of necrotic tissue, and control of infection and inflammation, but recently the use of growth factors has been approved to accelerate the healing process and heal the wound. Human recombinant growth factor PDGF-BB is one of the first types of recombinant growth factors approved in treating diabetic wounds. Several studies have reported that PDGF is an important mediator in wound healing that helps to accelerate healing, improve inflammation, cell proliferation, angiogenesis, and tissue regeneration. In this study, the human PDGF-B gene sequence was inserted into the pET 21a (+) expression vector for cloning and then inserted under the T7 promoter for its expression in the E. coli shuffle host. Purification was done using a nickel agarose column and to check the activity of the purified protein, cell proliferation, migration, and interaction were checked. The results of this study showed that the dimer type of PDGF expressed and purified in the bacterial host, probably due to maintaining the correct folded structure, has both main activities, i.e., cell proliferation due to the active binding to the cell receptor, as well as the ability to bind to fibrinogen.

Keywords

Wound healing

platelet-derived growth factor

Expression

Cloning

Subjects

Molecular biotechnology

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