Meenatchisundaram S, Parameswari G, Michael A, Ramalingam S
Department of Microbiology, PSG College of Arts and Science, Coimbatore, India.
Int Immunopharmacol. 2008 Aug;8(8):1067-73. doi: 10.1016/j.intimp.2008.03.017. Epub 2008 Apr 21.
Antivenom antibodies were raised in 24-week-old white leghorn chickens against hemotoxic venoms of Russell's viper and Saw-scaled viper snakes. Booster injections of increasing concentrations of venom were given at 14days of time interval to raise the antivenom level in egg yolk. Antibodies were extracted from immunized chicken egg yolk by Polson et al. (Polson A., Von Wechmar M.B., Van Regenmortel M.H.V. Isolation of viral IgY antibodies from yolks of immunized hens. Immunological Communications 1980; 9:475-493.) and further purified by DEAE cellulose ion exchange column chromatography, which gave pure (180-200kDa) specific antibodies against venom. High titre of more than 1:10,000 antibodies were detected by ELISA at the 135th day of observation. The lethal toxicity and various pharmacological activities like hemorrhagic activity, phospholipase activity, edema and procoagulant activities of venom were carried out by both in vivo and in vitro methods. The effectiveness of antivenom in neutralizing these effects was carried out involving pre-incubation type experiments. The median effective dose (ED50) for Russell's viper venom was 0.96mg/2LD50/18g mice and for Saw-scaled viper venom it was 1.28mg/2LD50/18g mice. One millilitre of specific antivenom was effective in neutralizing 0.110mg of Russell's viper and 0.137mg of Saw-scaled viper venoms respectively (PD50). Antivenom was effective in neutralization assays in a dose dependent manner. The results indicate that antibodies raised in chicken could effectively neutralize the pharmacological effects induced by venoms and chickens therefore present an alternative and cheaper source of specific antibody generation.
在24周龄的白来航鸡中制备抗蛇毒抗体,针对罗素蝰蛇和锯鳞蝰蛇的血液毒素毒液。每隔14天注射递增浓度的毒液进行加强免疫,以提高蛋黄中的抗蛇毒水平。通过Polson等人(Polson A., Von Wechmar M.B., Van Regenmortel M.H.V. Isolation of viral IgY antibodies from yolks of immunized hens. Immunological Communications 1980; 9:475 - 493.)的方法从免疫鸡蛋黄中提取抗体,并通过DEAE纤维素离子交换柱色谱进一步纯化,得到针对毒液的纯(180 - 200kDa)特异性抗体。在观察的第135天,通过ELISA检测到抗体滴度高于1:10,000。通过体内和体外方法对毒液的致死毒性以及各种药理活性,如出血活性、磷脂酶活性、水肿和促凝血活性进行了研究。通过预孵育类型实验研究了抗蛇毒血清中和这些作用的有效性。罗素蝰蛇毒液的半数有效剂量(ED50)为0.96mg/2LD50/18g小鼠,锯鳞蝰蛇毒液的半数有效剂量为1.28mg/2LD50/18g小鼠。1毫升特异性抗蛇毒血清分别能有效中和0.110mg的罗素蝰蛇毒液和0.137mg的锯鳞蝰蛇毒液(PD50)。抗蛇毒血清在中和试验中呈剂量依赖性有效。结果表明,鸡产生的抗体能够有效中和毒液诱导的药理作用,因此鸡是产生特异性抗体的一种替代且更廉价的来源。