体外激活人组织转谷氨酰胺酶的免疫测定。

Immunoassay of in vitro activated human tissue transglutaminase.

机构信息

Institute of Laboratory Medicine, University Hospital Leipzig, D-04103 Leipzig, Germany.

出版信息

Anal Biochem. 2011 Apr 1;411(1):10-5. doi: 10.1016/j.ab.2010.12.011. Epub 2010 Dec 10.

DOI:10.1016/j.ab.2010.12.011

Abstract

Tissue transglutaminase (tTG) is a calcium-dependent enzyme that exerts a variety of physiological functions and is involved in various pathoprocesses. To characterize the role of tTG in disease, simple assays are necessary for detection. We developed a highly sensitive enzyme-linked immunosorbent assay (ELISA) that combines a transamidation step with immunological detection to determine tTG. This assay is based on covalent binding of in vitro activated tTG to N,N'-dimethylated casein and subsequent detection of coupled tTG by specific immunoglobulin G (IgG) antibodies directed against tTG followed by binding of a secondary biotin-labeled antibody. The assay reaches a detection limit of 0.05ng of tTG/ml. Type 1 and 3 transglutaminases and factor XIII are not detected. By use of this assay, tTG in several cell lines and the stimulatory effect of retinoic acid on tTG expression could be demonstrated. The new assay represents a promising tool for the study of tTG in normal cell physiology and under pathological conditions.

摘要

组织转谷氨酰胺酶（tTG）是一种依赖于钙的酶，具有多种生理功能，并参与各种病理过程。为了研究 tTG 在疾病中的作用，需要简单的检测方法来进行检测。我们开发了一种高度敏感的酶联免疫吸附测定法（ELISA），该方法将转酰胺步骤与免疫学检测相结合，以检测 tTG。该测定法基于体外激活的 tTG 与 N,N'-二甲酰化酪蛋白的共价结合，随后通过针对 tTG 的特异性免疫球蛋白 G（IgG）抗体检测结合的 tTG，然后结合二级生物素标记的抗体。该测定法的检测限为 0.05ng tTG/ml。该测定法不检测 1 型和 3 型转谷氨酰胺酶以及因子 XIII。使用该测定法，可以证明几种细胞系中的 tTG 以及维甲酸对 tTG 表达的刺激作用。该新测定法代表了研究正常细胞生理学和病理条件下 tTG 的有前途的工具。