Objective        To purify specific polyclonal antibodies against Deinagkistrodon acutus venoms for new antivenom research.       Methods        New Zealand white rabbits were immunized with Deinagkistrodon acutus venom. The IgG fraction was purified by affinity chromatography with the Econo-Pac Protein A Kit. Specific antibodies were purified by immuno-affinity chromatography with CNBr activated sepharose 4B.Then the polyclonal antibodies were tested by ELISA and Western blotting to evaluate its immunocompetence. Antibody biological activity of the polyclonal antibodies was evaluated by lethality neutralization assay and neutralization of venom hemorrhagic activ

ity.       Results        Through this way

we successfully prepared specific polyclonal antibodies against Deinagkistrodon acutus venoms. ELISA showed the antibody titers higher than 3 200. Western blotting indicated that the polyclonal antibodies specifically recognized the most protein of Deinagkistrodon acutus venoms. Neutralization of venom hemorrhagic activity demonstrated the polyclonal antibodies effectively neutralized subcutaneous hemorrhage induced by Deinagkistrodon acutus venoms. In the lethality neutralization assay

the polyclone antibodies provided 100% and 54% protection against 2LD

50

and 4LD

50

dose Deinagkistrodon acutus venoms respectively.       Conclusion        The specific polyclonal antibodies against Deinagkistrodon acutus are purified successfully with good specificity and bio-activity.