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针对肾刷状缘膜麦芽糖酶的单克隆抗体:与年龄相关的抗原改变

Monoclonal antibodies to renal brush border membrane maltase: age-associated antigenic alterations.

作者信息

Reiss U, Sacktor B

出版信息

Proc Natl Acad Sci U S A. 1983 Jun;80(11):3255-9. doi: 10.1073/pnas.80.11.3255.

Abstract

Monoclonal antibodies to maltase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) from young adult and aged rats were prepared by the hybridoma technique. Four cell lines producing antibodies of the IgG1 subclass to maltase were established. Two, designated 1F12E1 and 8B1G6, produced monoclonal antibodies specific for the catalytically active form of the enzyme found predominantly in enzyme preparations from young animals. The other two clones designated 7G10H3 and 2E1C10 produced monoclonal antibodies that reacted exclusively with an enzymatically inactive form of maltase found mostly in enzyme preparations from aged rats. The increased prevalence of an inactive form of the enzyme in the old rat accounts for the decreased maltase-specific activity previously reported in the senescent rat. The active and inactive maltase species were separated by immunoaffinity chromatography by using the monoclonal antibodies as ligands. The separated forms of the enzyme were not distinguished by NaDodSO4/polyacrylamide gel electrophoresis, peptide mapping of the CNBr-cleaved proteins, and the NH2-terminal residues of these peptides. This study demonstrates the presence of an altered, antigenically distinct enzyme in senescent animals. Critical issues on the mechanism of the aging process may be addressed by application of these findings.

摘要

采用杂交瘤技术制备了针对幼年和老年大鼠麦芽糖酶(α-D-葡萄糖苷葡萄糖水解酶,EC 3.2.1.20)的单克隆抗体。建立了四个产生IgG1亚类麦芽糖酶抗体的细胞系。其中两个命名为1F12E1和8B1G6的细胞系产生的单克隆抗体对主要在幼年动物酶制剂中发现的酶的催化活性形式具有特异性。另外两个命名为7G10H3和2E1C10的克隆产生的单克隆抗体仅与主要在老年大鼠酶制剂中发现的无酶活性形式的麦芽糖酶发生反应。老年大鼠中该酶无活性形式的患病率增加,这解释了先前报道的衰老大鼠中麦芽糖酶特异性活性降低的原因。通过使用单克隆抗体作为配体的免疫亲和色谱法分离了活性和无活性的麦芽糖酶种类。通过NaDodSO4/聚丙烯酰胺凝胶电泳、CNBr裂解蛋白的肽图谱分析以及这些肽的NH2末端残基,无法区分分离出的酶的形式。这项研究证明了衰老动物中存在一种改变的、抗原性不同的酶。应用这些发现可能会解决衰老过程机制的关键问题。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e47/394019/6a8ad856059e/pnas00637-0122-a.jpg

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