Production of HTLV-1 Protease Recombinant Molecule Fused to Human IgG Fcγ1 (HTLV-1 Protease:hFcγ1) for Targeting HTLV-1-associated Diseases Treatment

Document Type : Original Article(s)

Authors

1 Blood Borne Infections Research Center, Academic Center for Education, Culture and Research (ACECR), Razavi Khorasan, Mashhad, Iran

2 Association Professor, Immunology Research Center, Inflammation and Inflammatory Diseases Division, Mashhad University of Medical Sciences, Mashhad, Iran

10.48305/jims.v43.i808.0224

Abstract

Background: Northeast Iran is recognized as an endemic region for HTLV-1 virus. Given the increasing prevalence of HTLV-1-associated diseases in the country, prevention and achieving effective treatment are essential. The aim of the present study was to design and produce the HTLV-1 protease fused to the Fcγ1 fragment of human antibody in the Pichia pastoris yeast expression system for its use in designing a viral protease inhibitor in future studies.
Methods: Following the design of the HTLV-1 protease:hFcγ1 gene construct, the optimized gene was inserted into the XhoI and NotI restriction sites of the pPICZαA expression vector. First, the recombinant plasmid was transformed into E. coli TOP10F' using the calcium chloride method and then electroporated into Pichia pastoris GS115. Recombinant clones containing the gene were selected and grown on zeocin-containing culture medium. Finally, SDS-PAGE and Western blotting were performed to confirm the expression of the recombinant protein.
Findings: In the present study, the design, cloning, and expression of the recombinant protein HTLV-1 protease:hFcγ1, utilizing the two elements HTLV-1 protease and human hFcγ1 molecule, were performed in the Pichia pastoris expression system. The recombinant protein HTLV-1 protease:hFcγ1 is a heavily glycosylated homodimer with a pI of 3.8 and an Mw of 50 kDa. The correct insertion of the recombinant construct into the pPICZαA expression vector was confirmed using AOX1 and α-factor primer pairs, amplifying fragments of 1388 bp and 1687 bp, respectively
Conclusion: The results of this research could provide a promising strategy for employing the recombinant protein HTLV-1 protease:hFcγ1 in designing suitable candidates for viral protease inhibitors and treating HTLV-1-associated diseases.

Highlights

Sanaz Ahmadi Ghezeldasht: Google Scholar, PubMed

Seyed Abdolrahim Rezaee: Google Scholar, PubMed

Arman Mosavat: Google Scholar, PubMed

Keywords

Main Subjects

References

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Journal of Isfahan Medical School
Volume 43, Issue 808
4th Week, May
May and June 2025
Pages 224-233

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History

  • Receive Date: 26 August 2024
  • Revise Date: 08 June 2025
  • Accept Date: 13 April 2025

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