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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。
Objective: To prepare and identify the rotavirus yolk immunoglobulin (IgY) .Methods: The human rotavirus Wa strain and the monkey rotavirus SA11 were prepared in batches by MA - 104 cell culture technology. SPF pure strains of Layian laying hens were boosted with purified virus + adjuvant, and specific immunoglobulins were prepared from chicken yolk by salting-out, column chromatography and ultrafiltration, and purified immunoglobulins were subjected to purification Biological activity research. Results: High titers of anti-rotavirus immunoglobulin were produced in the yolk of immunized chickens. The ELISA titers reached 1.5 × 10 5. Under the condition of regular immunization, the titers of yolk antibodies were maintained at high titers of 12 More than a month. The IgY antibody purified by defatted salting-out and chromatography showed that its immunological titer was stable for more than 6 months at 4 ℃ without significant decrease. Conclusion: The results of in vitro cytopathic effect test show that IgY antibody prepared in bulk has high titers of specific rotavirus neutralizing activity.