检验用大肠杆菌K88纤毛抗原的纯化及其单因子血清制备与质量控制方法研究

Purification of K88 Pilus Antigen of Escherichia coli,Preparation of Antiserum and Contrail of Process Quality for Testing

为研究兽用生物制品检验用大肠杆菌K88纤毛抗原的纯化及其单因子血清制备与质量控制方法。通过生物反应器发酵含K88抗原的大肠杆菌,机械脱纤处理获得K88纤毛抗原,经柠檬酸等电点沉淀和饱和硫酸铵梯度盐析以及透析处理纯化抗原,免疫家兔3次制备单因子血清并进行质量控制。结果显示,所纯化的K88纤毛抗原纯度能达到85%,每升发酵菌液可得到提纯抗原33.3 mg,免疫家兔血清琼扩效价可达到1∶128。表明该纤毛抗原制备和纯化及其单因子血清制备方法简易,纯度和产率较高,质量可控且易于批量制备。

In order to establish the method for purification of K88 pilus antigen of Escherichia coli,preparation of antiserum and contrail of quality of the process for testing of veterinary biological products.Escherichia coli of K88 pilus antigen was fermented by a bioreactor,K88 pilus antigen was broken off by mechanical method and purified by isoelectric precipitation of citric acid,gradient salting out of saturated ammonium sulfate and dialysis.Antiserum was prepared by immunizing rabbits 3 times and the quality of process was controlled.The results showed that the purity of K88 pilus antigen was about 85%,3.33 mg of K88 pilus antigen was produced by one liter of Escherichia coli.The AGP titer of its antiserum was 1∶128.The results suggest that the method for purifing of K88 pilus antigen and preparation of antiserum is relatively easy to be used to produce a large of mount of antigen and antiserum.The purity and productivity are relatively high,and the quality of process can be controlled.