A new luminescence assay for autoantibodies to mammalian cell-prepared insulinoma-associated protein 2.

A new luminescence assay for autoantibodies to mammalian cell-prepared insulinoma-associated protein 2.
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用于自身抗体的新发光测定法对哺乳动物预制作的胰岛素瘤相关蛋白2。

DOI:
10.2337/dc08-0286
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发表时间:
2008-09
期刊:
影响因子:
16.2
通讯作者:
Notkins, Abner Louis
Notkins, Abner Louis
中科院分区:
医学1区
文献类型:
--
作者:
Burbelo, Peter D.;Hirai, Hiroki;Leahy, Hannah;Lernmark, Ake;Ivarsson, Sten A.;Iadarola, Michael J.;Notkins, Abner Louis

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胰岛素瘤相关蛋白2(IA-2)是1型糖尿病的主要自身抗原,IA-2自身抗体通常通过液相放射免疫沉淀法检测。本实验的目的是开发一种新的测定方法,该方法不需要使用在细菌中制备的放射性同位素或自身抗原或通过体外转录/翻译制备。研究设计和方法-IA-2荧光素酶融合蛋白在哺乳动物细胞中表达,并通过液相荧光素酶免疫沉淀法测定自身抗体。结果-我们的研究表明,荧光素酶免疫沉淀法和放射免疫沉淀法在灵敏度和特异性方面没有显着差异,并且两种方法的比较显示出较高的相关系数(R2 = 0.805)。结论-荧光素酶系统提供了一个强大的,廉价的,非放射性的方法检测自身抗体的哺乳动物细胞制备的IA-2,并可能在临床水平上的实用价值。
OBJECTIVE—Insulinoma-associated protein 2 (IA-2) is a major autoantigen in type 1 diabetes, and IA-2 autoantibodies are routinely detected by a liquid-phase radioimmunoprecipitation assay. The present experiments were initiated to develop a new assay that does not require the use of radioisotopes or autoantigens prepared in bacteria or by in vitro transcription/translation. RESEARCH DESIGN AND METHODS—IA-2 luciferase fusion protein was expressed in mammalian cells and assayed for autoantibodies by liquid-phase luciferase immunoprecipitation. RESULTS—Our study showed that there was no significant difference between the luciferase immunoprecipitation and the radioimmunoprecipitation assays in sensitivity and specificity, and comparison of the two assays revealed a high correlation coefficient (R2 = 0.805). CONCLUSIONS—The luciferase system offers a robust, inexpensive, nonradioactive method for the detection of autoantibodies to mammalian cell–prepared IA-2 and could be of practical value at the clinical level.
DOI: 10.1373/49.6.916
发表时间: 2003-06-01
期刊: CLINICAL CHEMISTRY
影响因子: 9.3
作者:
Westerlund-Karlsson, A;Suonpää, K;Hinkkanen, AE
通讯作者: Hinkkanen, AE
DOI: 10.1016/j.bbrc.2006.11.140
发表时间: 2007-01-26
影响因子: 3.1
作者:
Burbelo, Peter D.;Ching, Kathryn H.;Kovacs, Joseph A.
通讯作者: Kovacs, Joseph A.
DOI: 10.1016/j.bbrc.2007.11.077
发表时间: 2008-02-01
影响因子: 3.1
作者:
Burbelo, Peter D.;Groot, Sandra;Iadarola, Michael J.
通讯作者: Iadarola, Michael J.
DOI: 10.1172/jci14257
发表时间: 2001-11-01
影响因子: 15.9
作者:
Notkins, AL;Lernmark, Å
通讯作者: Lernmark, Å
DOI: 10.1016/j.clim.2008.01.018
发表时间: 2008-06-01
影响因子: 8.6
作者:
Hirai, Hiroki;Miura, Junnosuke;Notkins, Abner L.
通讯作者: Notkins, Abner L.