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针对核糖体P蛋白的自身抗体:在酶联免疫吸附测定中使用多抗原肽作为包被剂

Autoantibodies directed against ribosomal P proteins: use of a multiple antigen peptide as the coating agent in ELISA.

作者信息

Caponi L, Pegoraro S, Di Bartolo V, Rovero P, Revoltella R, Bombardieri S

机构信息

Clinical Immunology Unit, University of Pisa, Italy.

出版信息

J Immunol Methods. 1995 Feb 27;179(2):193-202. doi: 10.1016/0022-1759(94)00285-5.

Abstract

Autoantibodies directed against the ribosomal proteins P0, P1 and P2 (P proteins) are specific for systemic lupus erythematosus (SLE) and there are some evidences that they could be related to the neuropsychiatric manifestations of the disease. In this study, a multiple antigen peptide (MAP) carrying four copies of the C-terminal peptide (13 residues) of the P2 protein, which is a common epitope of the three P proteins, was prepared for use in an ELISA assay. It was employed to detect antibodies directed against the ribosomal P proteins in 102 SLE patients and the results were compared with those obtained using immunoblotting (IB). With this new ELISA, antiribosomal P protein antibodies were found in 15/102 SLE sera. These results correlated well with the results of IB. Furthermore, we confirmed that naturally occurring antiribosomal P protein antibodies are directed mainly against the epitope containing the C-terminal sequence and shared by the three P proteins. MAP appears to be an excellent coating agent for ELISA assays designed to detect anti-P antibodies. Further experiments showed the superiority of MAP, compared to the free peptide, in the detection of weakly positive sera. This ELISA can also be used for the serological follow-up of SLE patients.

摘要

针对核糖体蛋白P0、P1和P2(P蛋白)的自身抗体对系统性红斑狼疮(SLE)具有特异性,并且有一些证据表明它们可能与该疾病的神经精神表现有关。在本研究中,制备了一种携带P2蛋白C末端肽(13个残基)四个拷贝的多抗原肽(MAP),该肽是三种P蛋白的共同表位,用于酶联免疫吸附测定(ELISA)。用其检测102例SLE患者中针对核糖体P蛋白的抗体,并将结果与使用免疫印迹法(IB)获得的结果进行比较。使用这种新的ELISA方法,在102份SLE血清中有15份检测到抗核糖体P蛋白抗体。这些结果与IB结果高度相关。此外,我们证实天然存在的抗核糖体P蛋白抗体主要针对包含C末端序列且为三种P蛋白所共有的表位。MAP似乎是用于检测抗P抗体的ELISA检测的优良包被剂。进一步的实验表明,与游离肽相比,MAP在检测弱阳性血清方面具有优势。这种ELISA也可用于SLE患者的血清学随访。

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