HEV (Hepatitis E Virus) recombinant proteins are engineered proteins derived from the HEV genome. Hepatitis E virus is a major cause of viral hepatitis, particularly in developing countries, and is transmitted primarily through contaminated water. Recombinant HEV proteins are vital for research, diagnostics, and vaccine development.
Structure and Function
The HEV genome encodes several proteins, with key recombinant targets including ORF2 (capsid protein), ORF3 (multifunctional regulatory protein), and non-structural proteins from ORF1.
The ORF2 protein is the primary structural protein forming the viral capsid, critical for viral assembly, and the main target for antibody responses.
ORF3 is involved in virus-host interactions and modulates host cell processes to favor viral replication.
Production
HEV recombinant proteins are typically produced in various expression systems, including E. coli, insect cells, yeast, or mammalian cells, depending on the need for post-translational modifications.
Expression vectors containing the HEV gene of interest are used to express the recombinant protein, which is then purified using techniques such as affinity chromatography (e.g., using His-tagged proteins).
Applications
Diagnostics: Recombinant HEV proteins, particularly ORF2, are used in ELISA kits and other serological assays to detect HEV-specific antibodies in patient sera, enabling diagnosis of Hepatitis E.
Vaccine Development: ORF2 recombinant proteins are used to develop subunit vaccines that can generate an immune response without the risks associated with live virus vaccines.
Research: These proteins are essential in studying the immune response to HEV, virus-host interactions, and for screening potential antiviral drugs targeting HEV.
Validation and Quality Control
Recombinant HEV proteins undergo rigorous validation to ensure correct folding, biological activity, and purity, using methods like SDS-PAGE, Western blotting, and functional assays.
The functionality of these proteins is confirmed by their ability to bind to specific antibodies or to mimic the viral protein's natural function in cell-based assays
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