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人唾液α-淀粉酶特异性单克隆抗体的制备

Preparation of human salivary alpha-amylase specific monoclonal antibody.

作者信息

Ito K, Katayama M, Minamiura N, Yamamoto T

出版信息

J Biochem. 1985 May;97(5):1357-62. doi: 10.1093/oxfordjournals.jbchem.a135188.

Abstract

A mouse hybridoma cell line which produced an anti-human salivary alpha-amylase monoclonal antibody was obtained by fusion between mouse spleen cells immunized with human salivary alpha-amylase and mouse myeloma cells, followed by screening the hybridoma cells by enzyme-linked immunosorbent assay. The hybridoma cell line (27-4-1) secreted IgG. The monoclonal antibody produced by the hybridoma showed no inhibitory effect on the activity of human salivary alpha-amylase. The specificity and reactivity of this monoclonal antibody were examined by determining the activities of human salivary and pancreatic alpha-amylases bound to the monoclonal antibody immobilized on polystyrene balls or by enzyme immunoassay with the monoclonal antibody conjugated with beta-D-galactosidase. The results revealed that the monoclonal antibody produced by the hybridoma cell line was specific for salivary alpha-amylase and absolutely unreactive to pancreatic alpha-amylase.

摘要

用人唾液α-淀粉酶免疫的小鼠脾细胞与小鼠骨髓瘤细胞融合,然后通过酶联免疫吸附测定筛选杂交瘤细胞,获得了产生抗人唾液α-淀粉酶单克隆抗体的小鼠杂交瘤细胞系。该杂交瘤细胞系(27-4-1)分泌IgG。该杂交瘤产生的单克隆抗体对人唾液α-淀粉酶的活性无抑制作用。通过测定固定在聚苯乙烯球上的单克隆抗体结合的人唾液和胰腺α-淀粉酶的活性,或用与β-D-半乳糖苷酶偶联的单克隆抗体进行酶免疫测定,检测该单克隆抗体的特异性和反应性。结果显示,该杂交瘤细胞系产生的单克隆抗体对唾液α-淀粉酶具有特异性,对胰腺α-淀粉酶完全无反应。

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