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Published on: June 26, 2014
Recombinant multiepitope proteins for hepatitis C virus diagnosis: Systematic review
Raissa Martins Deodato1, Mayara Torquato Lima da Silva2, Eduarda Oliveira Queiroz1
1Instituto Oswaldo Cruz (IOC), Rio de Janeiro, Brazil.
Insights
Recombinant multiepitope proteins offer a cost-effective and scalable solution for Hepatitis C virus (HCV) diagnosis, showing high sensitivity and specificity. This approach enhances global HCV detection efforts, aiding in the WHO
Area of Science:
- Virology
- Immunology
- Biotechnology
Background:
- Hepatitis C virus (HCV) infection affects over 50 million people globally, with diagnosis hindered by cost and accessibility, especially in low-resource settings.
- Conventional serological assays for HCV antibodies face limitations due to high production costs and restricted accessibility.
- Recombinant multiepitope proteins present a promising alternative by combining conserved antigenic regions from multiple HCV proteins (Core, NS3, NS4, NS5) into a single construct.
Purpose of the Study:
- To systematically review studies on the design, expression, purification, and validation of multiepitope constructs for Hepatitis C virus (HCV) serodiagnosis.
- To assess the diagnostic performance of multiepitope antigens compared to conventional methods and commercial assays.
Main Methods:
- Systematic literature search conducted in PubMed, Scopus, LILACS, and ISI Web of Science databases, identifying ten eligible articles.
- Analysis of protein expression (primarily in Escherichia coli using pET-based vectors) and purification methods (affinity or ion-exchange chromatography).
- Evaluation of diagnostic platforms including ELISA, immunoblot, dot blot, and rapid lateral-flow assays using human serum or plasma samples.
Main Results:
- Multiepitope antigens demonstrated high diagnostic performance, with sensitivity ranging from 91% to 100% and specificity from 95% to 100%.
- Performance of multiepitope antigen-based assays often equals or surpasses that of commercial assays.
- Key factors for improved performance include strategic selection of conserved epitopes, computational epitope prediction, and the use of flexible linkers.
Conclusions:
- Recombinant multiepitope proteins offer an efficient, low-cost, and scalable diagnostic strategy for HCV.
- This approach significantly enhances global HCV detection capabilities, contributing to the World Health Organization's (WHO) 2030 elimination goals.
- Further multicenter validation and standardized production workflows are necessary to fully realize the potential of this diagnostic strategy.
Abstract:
Hepatitis C virus (HCV) infection remains as major global health concern given the more than 50 million chronic cases recorded and the significant barriers to its early diagnosis, mainly in low-resource settings. Conventional serological assays rely on recombinant or synthetic antigens accounting for higher production costs and for limiting diagnostic accessibility. Recombinant multiepitope proteins have emerged as promising alternative for incorporating conserved antigenic regions from multiple HCV proteins, such as Core, NS3, NS4 and NS5, into a single construct capable of broadly detecting anti-HCV antibodies across different genotypes. The aim of this systematic review is to assess studies that have designed, expressed, purified and validated multiepitope constructs for HCV serodiagnosis. A search was made in PubMed, Scopus, LILACS and ISI Web of Science databases and it led to ten eligible articles. Most proteins were expressed in Escherichia coli by using pET-based vectors; they were first purified through affinity or ion-exchange chromatography. Diagnostic assessments employed ELISA, immunoblot, dot blot and rapid lateral-flow platforms, and used human serum or plasma samples provided by different geographical regions. Multiepitope antigens recorded high diagnostic performance across studies: sensitivity ranged from 91% to 100% and specificity ranged from 95% to 100%. These numbers are often comparable to, or surpass, commercial assays. The strategic conserved epitopes and rational design approach selection, including computational epitope prediction and flexible linker insertion, were key elements to improve antigenicity and assay robustness. Despite the promising results, further multicenter validation and standardized production workflows are still needed. Overall, recombinant multiepitope proteins represented an efficient, low-cost and scalable diagnostic strategy assumingly enhancing global HCV detection efforts and supporting progress toward WHO's 2030 elimination goals.
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