抗日本血吸虫Sjp40鸡卵黄抗体的制备及其生物学特性鉴定

目的制备抗日本血吸虫蛋白（sjp40）鸡卵黄抗体（eggyolkimmunoglobulinY，IgY），以期为日本血吸虫病的免疫学诊断提供一种特异性强、灵敏度高的优势抗体。方法以纯化的sjp40抗原免疫健康SPF级产蛋鸡，初次免疫2周后开始每日收集的鸡蛋为样本，采用水稀释硫酸铵沉淀法从鸡蛋中初步提取抗sjp40IgY，进一步经透析袋、DEAE-FF阴离子交换柱纯化IgY。用SDS—PAGE、WesternBlot及ELISA对纯化后的抗sjp40IgY进行分析鉴定。结果纯化的特异性抗Sjp40IgY，抗体效价1：10。。相对分子质量为180kDa，SDS．PAGE可见有2条主要蛋白带，分别约为66kDa和35kDa。并可被sj040特异性识别。结论成功制备具有良好免疫反应性、特异性强、灵敏度高的抗Sip40特异性IgY抗体。

PREPARATION AND BIOLOGICAL CHARACTERIZATION OF EGG YOLK IMMUNOGLOBULIN Y AGAINST SCHISTOSOMA JAPONICUM PROTEIN 40

Objective To prepare and characterize chicken egg yolk immunoglobulin Y （IgY） against Schistosoma japonicum protein 40 （Sjp40） which is one of the potential candidates in early diagnosis of schistoso- miasis japonica, so as to provide a highly specific and sensitive antibody against Sip40 for immunological diagnosis of Schistosomiasis japonica. Methods The healthy SPF hens were immunized with purified Sjp40. Two weeks later, the eggs were collected, from which specific IgY against Sjp40 were purified using water extraction, ammonium sul- fate precipitation and then followed by DEAE-FF anion exchange column. Then the purified specific IgY against Sjp40 was characterized by polyacrylamide gel electrophoresis, Western blot and ELISA. Results The anti-Sjp40 IgY was about 180 kDa with two bands（65 kDa, 28 kDa）. The purified anti-Sjp40 IgY could specifically recognize Sjp40 with a titer of 1 ： 106. Conclusion Specific IgY against Sjp40 with good immunoreactivity was successfully prepared.