Cyclosporin A and its analogs inhibit hepatitis B virus entry into cultured hepatocytes through targeting a membrane transporter, sodium taurocholate cotransporting polypeptide (NTCP)

Watashi, Koichi; Sluder, Ann; Daito, Takuji; Matsunaga, Satoko; Ryo, Akihide; Nagamori, Shushi; Iwamoto, Masashi; Nakajima, Syo; Tsukuda, Senko; Borroto-Esoda, Katyna; Sugiyama, Masaya; Tanaka, Yasuhito; Kanai, Yoshikatsu; Kusuhara, Hiroyuki; Mizokami, Masashi; Wakita, Takaji

To quantify HBs protein by ELISA, microwell antigen capture plates (Maxisorp nunc-immuno plate, Nunc #439454) were prepared by overnight incubation at 4ËšC with a sheep anti-HBs antibody at 1:5000 dilution, followed by coating with 0.2% BSA/0.02% NaN 3 /1 x PBS at 4ËšC until use. For HBs detection, samples were incubated in individual wells of the capture plates for 2 h. After washing, horseradish peroxidase-labeled rabbit anti-HBs antibody was added for an additional 2 h incubation. The substrate solution (from the HCV core ELISA kit: Ortho) was reacted for 15-60 min before the OD 450 values were measured.