Telakowski-Hopkins C A, Lu A Y, Pickett C B
Arch Biochem Biophys. 1983 Feb 15;221(1):79-88. doi: 10.1016/0003-9861(83)90123-6.
We have established hybridoma cell lines which secrete monoclonal antibody to rat liver microsomal epoxide hydrolase. All of the monoclonal antibodies formed against epoxide hydrolase are mouse immunoglobulin subclass IgG1. Utilizing double immunodiffusion analysis, we have found that the monoclonal antibodies bind to and precipitate epoxide hydrolase in solubilized rat liver microsomes. Despite the ability of the monoclonal antibodies to precipitate rat liver microsomal epoxide hydrolase, they do not inhibit the catalytic activity of the enzyme but rather stimulate it. The monoclonal antibodies do not precipitate epoxide hydrolase in microsomes isolated from hamster, rabbit, monkey, or human. When rabbit reticulocyte lysates are programmed with rat liver mRNA, the primary translation product of epoxide hydrolase can be immunoprecipitated from the translation system using the monoclonal antibodies. The ability of the antibodies to recognize in vitro synthesized epoxide hydrolase should make them amenable to identify recombinant bacterial clones expressing this protein. The monoclonality of these antibodies and their specificity should provide a useful way of identifying, purifying, and quantitating rat liver epoxide hydrolase as well as examine the expression of the gene(s) encoding epoxide hydrolase in normal rats and in rats exposed to carcinogenic xenobiotics.
我们已经建立了能分泌抗大鼠肝微粒体环氧化物水解酶单克隆抗体的杂交瘤细胞系。所有针对环氧化物水解酶形成的单克隆抗体均为小鼠免疫球蛋白亚类IgG1。利用双向免疫扩散分析，我们发现这些单克隆抗体能与溶解的大鼠肝微粒体中的环氧化物水解酶结合并使其沉淀。尽管这些单克隆抗体能够沉淀大鼠肝微粒体环氧化物水解酶，但它们并不抑制该酶的催化活性，反而会刺激其活性。这些单克隆抗体不能沉淀从仓鼠、兔子、猴子或人类分离的微粒体中的环氧化物水解酶。当用大鼠肝mRNA对兔网织红细胞裂解物进行编程时，环氧化物水解酶的初级翻译产物可以使用单克隆抗体从翻译系统中免疫沉淀出来。抗体识别体外合成的环氧化物水解酶的能力应使其适合于鉴定表达该蛋白的重组细菌克隆。这些抗体的单克隆性及其特异性应为鉴定、纯化和定量大鼠肝环氧化物水解酶以及检测正常大鼠和暴露于致癌性异源生物的大鼠中环氧化物水解酶编码基因的表达提供一种有用的方法。
