单克隆抗体在鸡传染性法氏囊病上的应用

THE APPLICATION OF McAb IN INFECTIOUS BURSAL DISEASE OF CHICKENS

本文将纯化的鸡法氏囊病毒免疫BALB/C小白鼠后,用免疫脾细胞与骨髓瘤细胞SP_2/O在PEG存在下进行融合,经HAT筛选,并通过3次克隆化后,获得3株能稳定分泌特异性单抗的杂交瘤细胞株。经ELISA测定,均属于IgG_1亚类。通过阻断试验证明其作用的特异性,并证明其在pH6～8下1小时,37℃时2天、-30℃下3个月均对效价无明显影响,而在60℃下30分钟,效价下降。应用特异性单抗作为诊断试剂来检测鸡法氏囊病是一种快速、灵敏、准确而有效的方法,仅需极微量的抗原(1μg/ml)即可显示阳性反应,不同品种发病鸡(三黄鸡、罗斯鸡、AA鸡)口服单抗一次,均有明显的疗效;发病鸡越早服用单抗、效果越好。初步建立了单抗制剂的检测体系及治疗方法、应用于BD的防治上。

Three stable hybridoma cell lines which secreted specific monoclonal antibodies have been obtained from the fussion of routine mycloma cells and spleen cells of BALB/C mice immunized with purified IBDV of chickens. After being selected by HAT, the hybridoma cells have been cloned three times, and the McAb of the three hybridoma cell lines finally obtained were all of IgG_1 isotype by ELISA test. The blocking test has proved their specificity, the titcrs of McAb were not affected at 37℃, two days; pH 6-8,1h; or-30℃, three months, but did come down at 60℃, 30 minutes. It's s quick, sensitive, correct and effective way to use specific McAbs as a diagnostic reagent to detect IBDV, which needs only a small amount of antigen to give the positive reaction. Sick chickens of different breeds could be cured by taking McAbs only one time, and the sick chickens took the McAbs earlier, the effect would be better. By the use of McAbs preparation, a detective and remedy system has been set up to apply to the prevention and cure of IBD.