抗黄曲霉尿酸氧化酶单克隆抗体的制备和初步鉴定

目的:制备抗黄曲霉尿酸氧化酶单克隆抗体。方法:采用杂交瘤技术,获得2株针对重组黄曲霉尿酸氧化酶(rUOX)的杂交瘤细胞McAb17和McAb3;采用盐析和A蛋白亲合层析柱纯化该抗体。结果:McAb17和McAb3的腹水效价达到1∶512000,纯化后获得纯度大于95%的单抗,抗体亚类(型)分别为IgG1/k型和IgG2a/k型;ELISA结果显示制备的单抗与rUOX和Rasburicase(商品化rUOX)可发生特异反应。结论:制备了抗黄曲霉尿酸氧化酶单克隆抗体McAb17和McAb3,为检测rUOX在动物体内的代谢变化提供了良好的实验基础。

Preparation and Identification of Monoclonal Antibodies Against Recombinant Urate Oxisase of Aapergillus flavus

Objective:To prepare the monoclonal antibodies(McAbs)against recombinant urate oxisase of Aapergillus flavus.Method:McAbs against recombinant urate oxidase(rUOX)of A.flavus were produced by fusing SP2/0 myeloma cells with spleen cells from BALB/c mice immunized with the purified rUOX.Two hybridoma cells secreting specific antibodies were obtained by screening culture supernatants by use of ELISA with the rUOX coated on the microplates.Results:One of the McAbs was IgG1/k,the other was IgG2a/k.The titers of the McAbs were up to 1:512 000.After two steps purification of salting-out and affinity chromatography,the McAbs with the purifity up to 95% were obtained.Both the purified McAbs can react with rUOX and the Rasburicase(commercialized rUOX of A.flavus).Conclusion:Monoclonal antibodies against recombinant urate oxidase(rUOX)of A.flavus were prepared,which lay a good experimental foundation for detecting metabolism of rUOX of animals in vivo.