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Submitted: 31 Aug 2019
Accepted: 22 Dec 2020
ePublished: 04 Jul 2020
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Avicenna J Pharm Res. 2020;1(1): 15-23.
doi: 10.34172/ajpr.2020.04
  Abstract View: 668
  PDF Download: 457

Research Article

Computational Design for the Production of Secretory Recombinant Codon-Optimized Human Stem Cell Factor (SCF) in Chinese Hamster Ovary (CHO) Cells With an Appropriate Signal Peptide: An Intensive In Silico Study

Ehsan Heidari Soureshjani 1,2*, Maryam Peymani 3*, Kamran Ghaedi 4,5

1 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
2 Shahrekord Regional Blood Transfusion Center, Shahrekord, Iran.
3 Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran.
4 Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran.
5 Department of Cellular Biotechnology, Cell Science Research Center, ACECR, Royan Institute for Biotechnology, Isfahan, Iran.
*Corresponding Authors: *Corresponding authors: Ehsan Heidari Soureshjani, MSc; SaNa Zist Pardaz Co, Member of Chahar Mahal and Bakhtiari Science and Technology Park, Shahrekord, Iran. Tel: +98-38-3226- 3854. Email: info@sanazist. ir, Maryam Peymani, PhD; Assistant Professor of Genetics, Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran. Tel: +98-38-3336-1010, Email: info@sanazist. ir; Email:

Abstract

Background: Production of the recombinant proteins in mammalian cells is an important issue with a bio-therapeutic purpose. Numerous efforts have been focused on the improvement of the yields of recombinant proteins, which include optimization of conventional biological processes, selection of appropriate signal peptides, codon optimization, and re-engineering of cells to produce more proteins. Stem cell factor (SCF) is a blood cytokine which activates the c-Kit receptor. This factor is crucial not only for the differentiation of hematopoietic progenitor cells but also for the survival, proliferation, and differentiation of mast cells. Recently, its therapeutic role in several diseases such as Alzheimer’s and myocardial infarction has been investigated. Therefore, the aim of this study was to design a secretory recombinant human SCF with the maximal yield in an appropriate mammalian host cell as Chinese hamster ovary (CHO) cells using the computational studies.

Methods: As the first step, computational simulation studies were carried out to design the appropriate signal peptide for the human SCF protein. Codon optimized coding sequence of hSCF was transferred into a eukaryotic expression vector (pBudCE4.1). Recombinant vector (pBudCE4.1/SCF) was transfected into CHO cells and the stably transformed cells were screened and isolated. Subsequently, the expression of SCF was defined by reverse transcription polymerase chain reaction (RT-qPCR) in stably transformed cells.

Results: Our bioinformatics studies indicated that Azurocidin signal peptide could be a suitable signal peptide for the production of SCF proteins in the CHO cells. Accordingly, computational studies revealed that the presence of 6×His-tag did not have a significant impact on the three-dimensional structure of the protein. Furthermore, the expression of hSCF was significant in the stable CHO cells.

Conclusion: The use of this approach may, therefore, lead to the production of highly efficient recombinant hSCF, which would be feasible for the mass production of this factor for therapeutic purposes.

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