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基于单克隆抗体的酶联免疫吸附测定法检测多胺精胺

Assay of the polyamine spermine by a monoclonal antibody-based ELISA.

作者信息

Garthwaite I, Stead A D, Rider C C

机构信息

Department of Biochemistry, University of London, Egham, Surrey, UK.

出版信息

J Immunol Methods. 1993 Jun 18;162(2):175-8. doi: 10.1016/0022-1759(93)90382-h.

Abstract

Spermine-specific monoclonal antibodies were prepared by immunising mice with protein-spermine conjugates. A resulting monoclonal antibody, IAG-1, exhibited both high affinity for spermine (binding constant 5.5 x 10(7) M-1) and high specificity, cross-reacting only weakly with spermidine. Using this antibody a competitive ELISA was developed with a detection limit of 1 pmol. The assay has been used to quantify spermine content of plant tissues, without derivatization, and producing within 6 h of collection, values for the spermine content which are similar to published data obtained by HPLC.

摘要

通过用蛋白质-精胺缀合物免疫小鼠制备了精胺特异性单克隆抗体。产生的一种单克隆抗体IAG-1对精胺表现出高亲和力(结合常数5.5×10⁷ M⁻¹)和高特异性,仅与亚精胺有微弱的交叉反应。使用该抗体开发了一种竞争性ELISA,检测限为1 pmol。该测定法已用于定量植物组织中的精胺含量,无需衍生化,并且在采集后6小时内得出的精胺含量值与通过高效液相色谱法获得的已发表数据相似。

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