Monoclonal Antibody Production against Yellow Fever NS1

Procedures to produce mAb against NS1 of YF were followed the methods described earlier [16 (link),17 (link)] with some modifications. Briefly, a 6-week SPF female mouse (Samtaco, Suwon, Korea) was immunized primarily with yellow fever NS1 25 μg in 25 μl PBS mixed with Freund’s Complete adjuvant (Sigma Chem Co., St. Louis, Missouri, USA) of 1:1 volume. Two weeks later, secondary immunization was carried out using incomplete adjuvant (Sigma) by the same method. Two weeks later, antibody titration was identified through indirect ELISA with blood collected from the tail. An absorbance of 1: 1,000 diluents of 0.2 or higher seemed successful immunization, and 25 μg of YF NS1 50 μl was injected into the mice’s feet 5 days before cell fusion to boost the immunity.