摘要
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。
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 supematants by use of ELISA with the rUOX coated on the microplates. Resuits： 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.
出处
《生物技术通讯》
CAS
2007年第3期465-466,共2页
Letters in Biotechnology
