商品化小鼠单抗试剂中普遍添加了高浓度的保护蛋白,如牛血清白蛋白(BSA),这给抗体的固定化带来了严重的干扰。本研究基于超滤技术发展了一种超微量抗体试剂(用量为10μ,约含2μg抗体、100μg BSA)预处理方法,能有效去除商品化抗体试剂中的高浓度BSA,使微珠表面固定化小鼠单抗的数量显著提高,并在蛋白质检测中得到了较好的结果,为缓解抗体产品供应现状对蛋白质芯片研究工作的制约提供了一条简单实用的途径。
Experiments showed that the immobilization was influenced obviously by protective proteins such as bovine serum albumin (BSA) , which are commonly added at a high concentration to prevent the commercial mouse monoclonal antibody products from titer decrease. In order to improve the immobilization of these antibodies, a simple pretreatment protocol based on ultra-filtration was carried out to remove most BSA from ultra trace antibody, in which the dose of antibody product was only 10μL ( contain 2 μg antibody and 100μg BSA). The pretreatment process was proved to be efficient, and the antibody beads showed fine activity in protein detection. This pretreatment can be used in the lab research of protein chips, which requires for small amount but several kinds of antibodies, to ease the difficulty of finding appropriate antibody products.