Journal: Molecular Therapy. Methods & Clinical Development

Article Title: HBVZ10, an AAV8 vector-based new HBV therapy candidate for cccDNA elimination

doi: 10.1016/j.omtm.2025.101646

Figure Lengend Snippet: Progressive reduction in serum HBsAg detected with ELISA was corroborated with western blot results and supported by intracellular HBsAg reduction (A) Progressive reduction of serum HBsAg detected by ELISA in mouse 838 (A1 orange) who received a single dose of HBVZ10 combined with 12-week ETV and was corroborated by western blot (A2). Mouse 831 was a mock-treated control. 100 × Hep: Positive control with concentrated viral particles from the HepAD38 cell medium. (B) Progressive reduction of serum HBsAg detected by ELISA in mouse 970 (B1 orange) who received mouse anti-HBs antibody monotherapy at a dose of 250 μg/injection triweekly for 9 consecutive times and was corroborated by western blot (B2). Mouse 907 was a mock-treated control. ADR and ADW: HBV positive human sera. 100xHepAD, Positive control with concentrated viral particles from HepAD38 cell medium. 35526 and 34477, negative human sera. X, no sample lane. Intrahepatic HBsAg was undetectable by ELISA (C1) in lysates (30 μL each) from seven mice that achieved an HBsAg − /anti-HBs + serum status. Consistently, western blot analysis (C2) of 40 μL liver lysates showed either undetectable or markedly reduced HBsAg levels in these seven mice compared with three untreated controls (mice 907, 987, and 471). (D) Intrahepatic HBc protein analyzed using western blot. 100xHepAD, Positive control with concentrated viral particles from HepAD38 cell medium. 100× GFP-HepG2 cell lysate, negative control. BHK-020, Uninfected liver lysate as negative control. X, no sample lane. Mice 812, 813, 814, 824, and 838 in green were the five mice with progressive reduction of serum HBsAg level upon blocking cccDNA replenishment. Mice 831 and 805 in blue were mock treated. Mouse 833 in purple received a single dose of HBVZ10 monotherapy on day 44pi. Four liver lysates from each liver were analyzed, and numbered as 1, 2, 3, and 4 or 5, 6, 7, and 8 or 11, 12, 13, and 14, respectively after mouse ID number.

Article Snippet: Serum anti-HBs antibody levels were quantified using the anti-HBs antibody ELISA kit (MONOLISA Anti-HBs EIA, cat. no. 25220, Bio-Rad) with corresponding calibrators (cat. no. 25219, Bio-Rad), following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Control, Positive Control, Injection, Negative Control, Blocking Assay