Double antibody sandwich ELISA was established with the recombinant antibodies against VZV glycoprotein and used for the determination of VZV glycoprotein level. The antibody combination， antibody concentration， coating conditions， blocking conditions， enzyme-labled antibody reaction conditions，and chromogenic condition were optimizated in the experiment， and the established method was validated for the specificity， linearity， accuracy， and precision.

The optimal combination of the double antibody was the group of VZV-mAb-03 as the capture antibody at a concentration of 450.0 μg·L^－1 and VZV-mAb-HRP-02 as the enzyme-labeled antibody at a concentration of 0.1 μg·L^－1； the optimal buffer for antibody coating was 0.05 mol·L^－1 Tris-HCl（pH=8.7±0.1），while the optimal temperature and time for coating were 4 ℃ and overnight（18 h） respectively； the bovine serum albumin（BSA） at a concentration of 1% was served as the blocking agent， while the optimal temperature and time for blocking were 37 ℃ and 2 h， respectively； the optimal reaction time of VZV and enzyme-labeled second antibody with PBST were both 1 h at the temperature of 37 ℃， and the optimal temperature and time for color reaction were 37 ℃ and 15 min， respectively. It was proved that this method had no cross reactions with the other vaccines or ingredients. The quantitation scope was 500－2 600 PFU·mL^－1， regression coefficient（R² ）> 0.95； the in-batch recovery rate was 82.2%－115.7% ， and the in-batch coefficient of variable（CV）< 10.2%； the batch-to-batch recovery was 80.5%－118.6%， the CV in validation for inplate<17.4% and the CV inter-plate precisions<14.9%.

Double antibody sandwich ELISA suitable for determination of VZV glycoprotein level is established，which meets the requirements for quantitative determination， and might be used to determine the VZV antigen level during the development and production of VZV vaccine.