The monoclonal antibody (mAb) D32.10 generated by HCV particles recognizes a discontinuous epitope encompassing three regions E1 (aa297-306),E2A(aa480-494) and E2B(aa613-621) juxtaposed on the surface of serum-derived HCV particles (HCVsp).To study the neutralizing capability of the E 1E2-specific D32.10 mAb,an in vitro direct cell-binding assay and an infection system of the human HepaRG cell line were developed by using HCVsp.The HepaRG cells possess potent ability to acquire a mature hepatocyte phenotype.The mAb D32.10 was shown to inhibit efficiently and specifically high affinity-interactions through glycosaminoglycans and the CD81 tetraspanin between HCVsp and human hepatocytes with an IC50 ≤ 0.5μg/ml.Otherwise,establishment of infection,replication and propagation of HCVsp were shown to depend on the proliferation/differentiation stage of HepaRG cells.Persistent HCV infection in HepaRG cells was obtained with production of E1E2/RNA(+) infectious HCV particles.Data showed a complete early inhibitory effect of D32.10 on virion RNA production in HepaRG culture supernatants.