Preventative Vaccines for Zika Virus Outbreak: Preliminary Evaluation

Kim, Eun; Erdos, Geza; Huang, Shaohua; Kenniston, Thomas; Falo, Louis D.; Gambotto, Andrea

Sera from the animals were collected every two weeks and tested for ZIKV-specific IgG by conventional ELISA. Briefly, ELISA plates were coated with 2 × 10 5 pfu of heat-inactivated ZIKV DAKAR4542 at 60°C for 20 min per well overnight at 4°C in carbonate coating buffer (100 mM, pH 9.5) and then blocked with PBS containing 0.05% Tween 20 (PBS-T) and 2% BSA for 1 h. Mouse sera were diluted 1:200 or 1:20 for pups sera in PBS-T with 1% BSA and incubated for 2 h. After the plates were washed, HRP-conjugated anti-mouse IgG (1:2000, Santacruz) was added to each well and incubated for 1 h. The plates were washed three times and developed with 3,3′5,5′-tetramethylbenzidine, and the reaction was stopped with 1 M H 2 SO 4 and absorbance at 450 nm was determined using an ELISA reader (BIO-TEK instruments).