Immunoassay of pig and human gastric proton pump.
Immunoassay of pig and human gastric proton pump.
复制标题
猪和人胃质子泵的免疫测定。
DOI：
10.1016/0016-5085(86)91105-4
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发表时间：
1986
期刊：
影响因子：
29.4
通讯作者：
Weinstein,WM
中科院分区：
文献类型：
--
作者：
Smolka,A;Weinstein,WM
通过杂交瘤技术制备了针对负责猪胃粘膜酸分泌的K+依赖性腺苷三磷酸酶（ATP酶）的单克隆抗体。其中两种抗体显示出选择性结合ATP酶亚基并标记猪和兔壁细胞的细胞内膜，用于开发ATP酶的灵敏（±1 pmol）免疫测定法。将酶样品吸附到聚苯乙烯微量滴定板的威尔斯孔中，然后依次与单克隆抗体、偶联碱性磷酸酶的抗小鼠免疫球蛋白G和对硝基苯基磷酸盐孵育。通过三参数逻辑拟合将威尔斯孔在410 nm处的吸光度与log 10 μ g ATP酶相关的标准曲线，有用的测定范围为0.05-10 μg ATP酶。免疫测定允许测量质子泵ATP酶水平在人类胃活检标本，因此可能是有用的胃粘膜功能的研究。
Monoclonal antibodies against the K+-dependent adenosine triphosphatase (ATPase) responsible for acid secretion in the pig gastric mucosa were generated by hybridoma technology. Two of these antibodies, shown to bind selectively to subunits of the ATPase and to label intracellular membranes of pig and rabbit parietal cells, were used to develop a sensitive (±1 pmol) immunoassay for the ATPase. Enzyme samples were adsorbed to the wells of polystyrene microtitration plates and then incubated sequentially with monoclonal antibody, antimouse immunoglobulin G coupled to alkaline phosphatase, and p-nitrophenyl phosphate. Standard curves relating the absorbance of the wells at 410 nm to log10micrograms ATPase were fitted by a three-parameter logistic, with a useful assay range of 0.05-10 μg ATPase. The immunoassay allows measurement of proton-pumping ATPase levels in human gastric biopsy specimens and may therefore be useful in studies of gastric mucosal function.