抗人血白蛋白单克隆抗体的制备及其免疫金标试纸条的研制

Establishment of a monoclonal antibody against human albumin and gold immunochromatography assay

目的:制备抗人血白蛋白单克隆抗体并对其抗体特性进行鉴定,将其标记胶体金,研制一种快速准确检测尿中人血白蛋白试纸条的方法。方法:用99.8%纯度的人血白蛋白免疫Balb/C小鼠,采用杂交瘤细胞技术制备单克隆抗体,并对其单克隆抗体的相关性质进行鉴定,制备胶体金,并标记所制备的单克隆抗体,按竞争法制备试纸条。结果:得到两株能够稳定分泌抗体,效价高的杂交瘤细胞系4C2,2F10。结论:建立的免疫层析试纸条,简便、可靠,有望通过进一步改进应用于临床检验。

Objective：To establish and identify of the antiAlb antibodies, Gold colloid was prepared and used to label a monoclonal antibody against human albumin. To develop a sensitive, rapid and simple gold immunochromatography assay for determination of the lever of human albumin in urine. Methods： Balb/c mice were immunized with protein and the immunized spleen cells were fused with SP2/0 to raise hybridoma. And identification of the antibodies, based on two monoclonal antibody （4C2,2F10） sandwich assay to determine human Albumin in the urine was developed. Gold colloid was prepared and used to label one of monoclonal antibody against human Albumin. Results： Two hybridoma cell lines named 4C2 and 2F10 were established, which could secrete high quality mAb. Conclusion：We have developed a sensitive, rapid and simple gold immunochromatography assay. It may have a broad prospect in clinical diagnosis.