抗轮状病毒卵黄免疫球蛋白制备工艺及生物学活性研究  

Preparation and identification of egg yolk immunoglobulin against rotavirus

作　　者：荫俊[1] 侯晓军[1] 张松乐[1] 王忠泽[1] 宋伟[1] 孙国章[1] 王汝懿 

机构地区：[1]中国人民解放军军事医学科学院微生物流行病研究所,北京100071

出　　处：《解放军药学学报》2000年第2期59-61,共3页Pharmaceutical Journal of Chinese People's Liberation Army

基　　金：国家新药基金!专题合同编号 :96 - 90 1 - 0 5 - 1 65

摘　　要：目的 :抗轮状病毒卵黄免疫球蛋白 (yolkimmunoglobulins,IgY)的制备及鉴定方法方法 :采用MA - 10 4细胞转瓶培养技术 ,批量制备人型轮状病毒Wa株和猴型轮状病毒SA11,以纯化病毒 +佐剂对SPF纯系莱航产蛋母鸡进行强化免疫 ,通过盐析、柱层析和超滤方法从鸡卵黄中制备特异性免疫球蛋白 ,并对纯化的免疫球蛋白进行生物学活性研究。结果 :免疫鸡卵黄中产生高滴度特异性抗轮状病毒免疫球蛋白 ,其ELISA免疫效价达 1.5× 10 5,在定期加强免疫条件下 ,卵黄抗体效价维持在高滴度可达 12个月以上。经脱脂盐析和层析纯化的IgY抗体 ,在 4℃条件下其免疫效价可稳定保存 6个月以上 ,无显著下降。结论 :体外细胞病变抑制实验结果表明 ,批量制备的IgY抗体具有高效价的特异性轮状病毒中和活性。Aim To study preparation and identification of egg yolk immunoglobulin against rotavirus.Methods With the cell roller culture of the MA-104 cells to batch culture of rotavirus Wa and SA 11 strains,SPF chickens were procedurally immunized with purified rotavirus.Special immunoglobulin against the rotavirus was prepared from immunized egg yolk with saltingout,column chromatography and ultra-filtration.Purified immunoglobulin was identified for stabilization and neutralization with the rotavirus.Results High titer egg yolk immunoglobulins(IgY) were produced by means of procedural immunization with purified rotavirus.The ELISA titer was measured at 1.5×10 5 and could be maintained at least 12 months by boosting immunization.Purified IgY was stable for 6 months in 4℃.Conclusion TCID 50 test revealed that prepared IgY could effectively suppress rotavirus pathogenic ability on the MA-104 cell in vitro.

关 键 词：卵黄 免疫球蛋白 轮状病毒 制备 生物学活性 

分 类 号：R392－33[医药卫生—免疫学]