Expression of the secrected NS1 of DENV2 virus and the preparation of the rabbit polyclonal anti-NS1 antibodies.

Objective To produce a secretary recombinant NS1 of dengue virus type 2 (DENV2) and prepare the anti-NS1 polyclonal antibody. Method The full-length NS1 gene was expressed with Pichia pastoris expression system and the recombinant NS-1 was used to immunize rabbit to prepare anti-NS1 polyclonal antibody. The antibody was detected by Western-blot and quantified by ELISA. The polyclonal antiserum was purified by caprylic acid-ammonium sulfate precipitation and protein A resin. The specificity and sensitivity of the antibody were determined by ELISA and Western-blot after purification. Result DENV2 NS1 protein was expressed efficiently in Pichia pastoris. The antibody could specifically recognize DENV2 NS1 protein and the titer of the antiserum obtained in this experiment was up to 1∶6000. Conclusion The successful preparation of the polyclonal antibody against NS1 protein provides an important reagent for further study of the role of NS1 in the pathogenesis of dengue virus.