一种新的皮肌炎自身抗体ELISA检测方法:将自身抗原cDNA快速引入重组法检测自身抗体。

Clinical & Developmental Immunology Pub Date : 2013-01-01 Epub Date: 2013-12-12 DOI:10.1155/2013/856815
Yoshinao Muro, Kazumitsu Sugiura, Masashi Akiyama
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引用次数: 32

摘要

免疫学、生物化学和分子生物学的进步使许多检测自身抗体的方法得以发展。ELISA之所以得到广泛应用,是因为它比免疫印迹和免疫沉淀等其他免疫方法能够更快地处理相对大量的血清样本。重组自身抗原通常是利用相关克隆cDNA在大肠杆菌中产生的,是ELISA所必需的。常规的临床酶联免疫吸附试验在纯化蛋白质不含细菌污染物的能力上是有限的,而且这个过程是劳动密集型的。我们最近开发了新的ELISA测试,利用自身抗原的简单体外转录和翻译标记来测量皮肌炎(DM)特异性自身抗体,包括Mi-2、MDA5、NXP-2、TIF1-α和TIF1-γ的自身抗体。这种方法可能允许将cdna快速转化为化学发光ELISA,以检测不仅在糖尿病而且在其他自身免疫性疾病中发现的自身抗体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A new ELISA for dermatomyositis autoantibodies: rapid introduction of autoantigen cDNA to recombinant assays for autoantibody measurement.

A new ELISA for dermatomyositis autoantibodies: rapid introduction of autoantigen cDNA to recombinant assays for autoantibody measurement.

A new ELISA for dermatomyositis autoantibodies: rapid introduction of autoantigen cDNA to recombinant assays for autoantibody measurement.

A new ELISA for dermatomyositis autoantibodies: rapid introduction of autoantigen cDNA to recombinant assays for autoantibody measurement.

Advances in immunology, biochemistry, and molecular biology have enabled the development of a number of assays for measuring autoantibodies. ELISA has been widely used, because it can deal with relatively large numbers of serum samples more quickly than other immunologic methods, such as immunoblotting and immunoprecipitation. Recombinant autoantigens, which are generally produced in E. coli using the relevant cloned cDNA, are necessary for ELISA. Conventional clinical ELISA tests are limited in their ability to purify proteins free of bacterial contaminants, and the process is labor intensive. We recently developed new ELISA tests that utilize simple in vitro transcription and translation labeling of autoantigens in order to measure dermatomyositis-(DM-) specific autoantibodies, including autoantibodies to Mi-2, MDA5, NXP-2, TIF1-α, and TIF1-γ. This method may allow for the rapid conversion of cDNAs to a chemiluminescent ELISA to detect autoantibodies that are found not only in DM but also in other autoimmune diseases.

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