副溶血性弧菌高免卵黄抗体的制备和不同提纯方法效果的比较

目的：通过比较聚乙二醇（PEG）沉淀法、氯仿抽提法和氯仿/PEG法3种卵黄抗体（IgY）提纯方法的提纯效果，为批量提纯高质量IgY提供依据。方法：制备副溶血性弧菌灭活疫苗，采用肌肉多点注射法免疫高产蛋鸡，收集鸡蛋，分别采用PEG沉淀法、氯仿抽提法和氯仿/PEG法对鸡蛋中的IgY进行提纯，比较3种提纯方法获得的IgY的蛋白质量浓度、抗体效价和纯度，结合操作过程、成本效益及安全性对3种提纯方法进行综合分析比较。结果：3种方法所提纯的IgY中，蛋白质量浓度从高到低依次为氯仿抽提法、氯仿/PEG法、PEG法；抗体效价相差不大，氯仿抽提法稍高；抗体纯度由高到低依次为PEG法、氯仿/PEG法、氯仿抽提法。PEG法安全性较好，但提取效率相对较低，成本投入较高；氯仿抽提法能够高效地从卵黄中获得IgY，但提取物中掺杂了大量的杂质蛋白；氯仿/PEG法提纯效率高，抗体纯度好，成本效益相对较高。结论：PEG法适用于实验室中少量IgY的提取；氯仿/PEG法的提取效率高，抗体纯度好，可用于批量提取高质量的IgY。

Preparation of high immunity yolk antibody against Vibrio parahemolyticus and comparison of effectiveness between different extraction methods

Objective:To compare the effectiveness between three methods for purifying the immunoglobulin of egg yolk(IgY) which are polyethylene glycol (PEG) method, chloroform extraction method and chloroform / PEG method, and to provide basis for obtaining the batch of IgY.Methods:The inactivated vaccine of Vibrio parahemolyticus (V. parahemolyticus) was prepared and the hens were immunized by multi-point intramuscular injection.The eggs were collected and the IgY was purified by PEG method, chloroform extraction method and chloroform/PEG method.The protein extraction rate, the IgY titer and the purity of the antibody which purified by different methods were detected.Furthermore, the operation process, cost and safety of the three methods were analyzed.Results:The protein contents of the extraction belonging three methods from high to low in turn were chloroform extraction method, chloroform/PEG method, and PEG method.There was no significant difference in the antibody titer between three methods, and the tiler of chloroform extraction method was slightly high.The purities of purified antibody from high to low in turn were PEG method, chloroform/PEG method and chloroform method.The PEG method had better security but relatively lower extraction efficiency and higher cost.The chloroform/PEG method had high extraction efficiency and good antibody purity.Conclusion:The PEG method is suitable for a small amount of extraction in the laboratory.The chloroform/PEG method is appropriate for extracting the high quality IgY in a batch as it has high extraction efficiency and good antibody purity.