Screening of anti-West Nile virus antibody drugs in vitro / 国际药学研究杂志

ABSTRACT

Objective: To screen anti-West Nile virus antibody drugs in vitro, using the previously established West Nile pseudovirus reporting system. Methods: At first, the enzyme-linked immunosorbent assay(ELISA)was used to detect the binding of 13 anti-West Nile virus antibody candidate strains previously obtained by the phage display technology to the target protein, West Nile virus E protein domainⅢ(DⅢ). Next, the collected cell culture supernatant containing the West Nile pseudovirus was diluted 102-fold, and the dilutions were incubated with the antibody candidates at relevant concentrations at room temperature for 1 h, respectively. Then, BHK21 or K562 cells were infected with the incubated pseudoviral supernatant+antibody mixture. After 48 hours, the infected cells were lysed to measure luciferase activity(RLU). Results: The tested 13 candidate antibodies could all recognize and bind to the West Nile virus E protein DⅢ. Among them, two antibodies(the antibodies 5 and 7)showed a good neutralizing activity and could effectively block the viral invasion upon target cells at the low concentration 5 mg/L, while two antibodies(the antibodies 3 and 8) showed an antibody-enhancing effect(ADE), which was manifested by enhancing viral infection of target cells. Conclusion: Using the West Nile pseudovirus reporting system that we established previously, we are able to obtain two anti-West Nile virus antibodies with neutralizing activity by the in vitro screening.