Expression, purification, structure and stability of recombinant bFGF from E.coli: A spectroscopic and calorimetry study

Ranjbar, Faezeh; Nasoohi, Nikoo; Khajeh, Khosro

doi:10.22034/NBR.12.2.2

Expression, purification, structure and stability of recombinant bFGF from E.coli: A spectroscopic and calorimetry study

نویسندگان

Faezeh Ranjbar ¹

Nikoo Nasoohi ²

Khosro Khajeh ³

¹ Department of Microbial Biotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.

² Department of Molecular and Cellular Sciences, Faculty of Advanced Sciences and Technology, Pharmaceutical Sciences Branch, Islamic Azad University (IAUPS), Tehran, Iran.

³ Department of Biochemistry, faculty of Biological sciences, Tarbiat Modares University, Tehran, Iran.

10.22034/NBR.12.2.2

چکیده

Basic fibroblast growth factor (bFGF), also known as FGF-2, is a crucial member of the fibroblast growth factor family, involved in a variety of biological functions including cellular proliferation, wound healing, angiogenesis, intercellular signalling, and cell differentiation, In contemporary stem cell research, serum-free media enriched with various additives and growth factors are employed, and among these, bFGF being particularly significant. Despite its extensive potential applications, the clinical utilization of bFGF is limited due to its instability, especially in aqueous environment. Therefore, a thorough investigation of the protein's structural integrity and stability is essential. This study focuses on the expression, purification, and characterization of bFGF for structural and stability analysis through biophysical methods. Intrinsic fluorescence measurement indicated a structural alteration surrounding the tryptophan residue, while circular dichroism (CD) analysis showed a decrease in the protein’s secondary structure. The differential scanning calorimetry (DSC) used for stability analysis. Furthermore, the study aims to evaluate the biological activity of the protein in cellular context. For this purpose, gold nanoparticles were synthesized. The results from the Cell Migration Assay indicated that the proliferation of HT29 cells was enhanced following treatment with bFGF in conjunction with gold nanoparticles. Also, a MTT assay was conducted.

کلیدواژه‌ها

Basic fibroblast growth factor

wound healing

Protein’s structure and stability- Biophysical approaches

fibroblast growth factor

purification

nanoparticle

20.1001.1.24236330.1404.12.2.3.2

موضوعات

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