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Article Abstract
Objectives: Development of an immunoassay for quantitation detecting the highly conserved hepatitis C core antigen (HCVcAg).
Methods: Anti-HCVcAg monoclonal antibodies (mAbs) were developed and evaluated using enzyme-linked immunosorbent assay (ELISA), and the chemiluminescence enzyme immunoassay (CLEIA) was also proposed. The serum HCVcAg was measured to evaluate and analyze the performance of CLEIA and analysis of the mAb pair was evaluated by sequencing of the antibody variable region.
Results: The prokaryotically expressed HCVcAg was purifed and three HCVcAg mAbs against HCVcAg with excellent detection performance were obtained after immunizing BALB/c mice. MAb 4D11 and 28B10 were selected as respective capture and detection antibodies for HCVcAg measurement by CLEIA (detection range, 0.1-256 μg/L). The results for CLEIA and fluorescent PCR used in hospitals demonstrated excellent consistency (K = 0.801, P < 0.01). The variable region genes of the heavy chain (VH) and light chain (VL) for the mAb pair were successfully sequenced.
Conclusion: The developed mAbs and CLEIA are expected to become a rapid and economical clinical diagnostic tool for HCVcAg detection.
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| http://dx.doi.org/10.1016/j.pep.2025.106747 | DOI Listing |
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