Comparing the Soluble Form of Recombinant Human Insulin-like Growth Factor-1 (rhIGF-1) in Using Thioredoxin As Fused and Co-expressed Protein

Overview

Journal Protein Pept Lett

Publisher Bentham Science Publishers

Specialty Biochemistry

Date 2024 Jul 4

PMID 38963110

Authors

Sara Hemmati

Parvaneh Maghami

Javad Ranjbari

Maryam Tabarzad

Affiliations

Soon will be listed here.

Abstract

Introduction: Insulin-like growth factor-1 (IGF-1) is a single-chain polypeptide with various physiological functions. is one of the most desirable hosts for recombinant protein production, especially for human proteins whose post-translation modifications are not essential for their bioactivity, such as hIGF-1.

Objectives: In this study, bacterial thioredoxin (Trx) was studied as a fused and non-fused protein to convert the insoluble form of recombinant human IGF-1 (rhIGF-1) to its soluble form in E. coli.

Methods: The rhIGF-1 was expressed in the Origami strain in the form of fused-Trx. It was co-expressed with Trx and then purified and quantified. In the next step, the biological activity of rhIGF-1 was evaluated by alkaline phosphatase (ALP) activity assay in human adipose- derived stem cells (hASCs) regarding the differentiation enhancement effect of IGF-1 through the osteogenic process.

Results: Results showed that Trx in both the fused and non-fused forms had a positive effect on the production of the soluble form of rhIGF-1. A significant increase in ALP activity in hASCs after rhIGF-1 treatment was observed, confirming protein bioactivity.

Conclusion: It was strongly suggested that the overproduction of Trx could increase the solubility of co-expressed recombinant proteins by changing the redox state in cells.

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