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Refolding of the recombinant protein OmpA70 from Leptospira interrogans from inclusion bodies using high hydrostatic pressure and partial characterization of its immunological properties

Paper ID Volume ID Publish Year Pages File Format Full-Text
24095 43496 2010 7 PDF Available
Title
Refolding of the recombinant protein OmpA70 from Leptospira interrogans from inclusion bodies using high hydrostatic pressure and partial characterization of its immunological properties
Abstract

Leptospira is the etiological agent of leptospirosis, a life-threatening disease that affects human populations worldwide. Available vaccines have demonstrated limited effectiveness, and therapeutic interventions are complicated by the difficulty of establishing an early diagnosis. The genome of Leptospira strains was sequenced, and bioinformatic analyses revealed potential vaccine and serodiagnosis candidates. The present work studied OmpA70, a putative outer membrane protein from Leptospira interrogans serovar Copenhageni that combines structural features of Loa22, the first genetically defined virulence factor in Leptospira, and Lp49, a protein that reacts with sera from early and convalescent patients. Recombinant OmpA was produced in Escherichia coli in an insoluble form. Considering the importance of the structural integrity of a protein to confer immune protection, high hydrostatic pressure (HHP) was used to refold OmpA70 aggregated as inclusion bodies. HHP was applied in association with redox-shuffling reagents (oxidized and reduced glutathione) and guanidine hydrochloride or l-arginine. About 40% of the protein was refolded by applying 200 MPa for 16 h in concentrations of l-arginine above 0.4 M. Circular dichroism revealed the presence of secondary structure. OmpA70 has immunogenic and antigenic properties as high antibody titers were seen after immunization with this protein, and sera from infected hamsters reacted with soluble OmpA70.

Keywords
HHP, high hydrostatic pressure; IBs, inclusion bodies; GdnHCl, guanidine hydrochloride; OmpA70, 70 kDa leptospiral protein with OmpA domain; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresisLeptospira; Refolding; High hydrostatic pressur
First Page Preview
Refolding of the recombinant protein OmpA70 from Leptospira interrogans from inclusion bodies using high hydrostatic pressure and partial characterization of its immunological properties
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 148, Issues 2–3, 20 July 2010, Pages 156–162
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering