PRODUCTION OF A MONOCLONAL-ANTIBODY AGAINST HUMAN AMELOGENIN
PRODUCTION OF A MONOCLONAL-ANTIBODY AGAINST HUMAN AMELOGENIN
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DOI：
10.1007/bf00316294
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发表时间：
1994-01-01
影响因子：
4.2
通讯作者：
DEUTSCH, D
中科院分区：
文献类型：
--
作者：
CATALANOSHERMAN, J;LASKOV, R;DEUTSCH, D
发育中的人类牙釉质的细胞外有机基质由两大类蛋白质组成，即疏水性的釉原蛋白和酸性的釉蛋白。为了从这种复杂的蛋白质混合物中鉴定、纯化和表征釉原蛋白，并研究它们的超微结构定位以及它们的合成、分泌和降解途径，需要特异性和灵敏的探针。在本文中，描述了采用脾内初次免疫方案生产抗人釉原蛋白的单克隆抗体。所产生的单克隆抗体为IgM，在蛋白质免疫印迹分析中可识别主要的人釉原蛋白条带。它还能识别其他物种（特别是牛和猪）的釉原蛋白条带。间接免疫组织化学研究表明，该单克隆抗体与人发育中的牙釉质细胞外基质特异性反应，而不与下方的牙本质层反应。
The extracellular organic matrix of developing human enamel is composed of two major classes of proteins, the hydrophobic amelogenins and the acidic enamelins. In order to identify, purify, and characterize the amelogenins from this complex mixture of proteins, and to study their ultrastructural localization and their pathways of synthesis, secretion, and degradation, specific and sensitive probes are needed. In the present paper the production of a monoclonal antibody against human amelogenin employing an intrasplenic primary immunization protocol is described. The monoclonal antibody produced is IgM and recognizes major human amelogenin protein bands in Western immunoblot assays. It also recognizes amelogenin protein bands from other species, specifically bovine and porcine. Indirect immunohistochemical studies showed the monoclonal antibody to react specifically with the extracellular matrix of human developing enamel. It did not react with the underlying dentin layer.