The invention discloses a method for refining a polyvalent ASV (anti-snake venom) lyophilized serum, pertaining to an immune serum containing antibody which aims at least two kinds of antigens. The serum takes the venom of Ancistrodon acutus, Agkistrodon halys, Pit-vipers, bamboo snakes, adders and the like as the immune antigen which is inoculated into the bodies of animals after degerming and dialysis, and then titer immune serum after purification is used for collecting the antibody which is then desalted and lyophilized; mice are used for evaluating the LD50 of the virulence of the venom and the titer of the ASV serum; the antibody inoculated into the bodies of the animals is one of the venom, toxoid or snake venom or the composition thereof, and glutaraldehyde or formaldehyde is used for toxin detoxication; before immunity, anti-tetanic toxoid condensed antibody for human beings is used for carrying out minim gradual immunization on the animals, or an Emuade adjuvant is used for immunization purpose. The polyvalent ASV lyophilized serum complies with the biological product standards upon the tests and inspections of purity and specific activity, safety, pyrogen, hypersusceptibility, dissolution and the like; moreover, the serum can cure sufferers who are bitten by vipers with the advantages of high purity, low foreign protein, easy preservation and quick dissolving capacity.