High-throughput screening for native autoantigen-autoantibody complexes using antibody microarrays.

High-throughput screening for native autoantigen-autoantibody complexes using antibody microarrays.
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DOI:
10.1021/pr4001674
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发表时间:
2013-05-03
影响因子:
4.4
通讯作者:
Lampe PD
Lampe PD
中科院分区:
生物学2区
文献类型:
--
作者:
Rho JH;Lampe PD

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我们报告了一种新的,高维的方法来检测自身抗体与其天然自身抗原复合。具体地,将血清或血浆中的自身抗体-自身抗原复合物直接温育到高密度抗体微阵列上。结合的自身抗体-抗原复合物的检测通过荧光标记的抗人免疫球蛋白G或其他免疫球蛋白同种型二抗进行,并在微阵列扫描仪中定量。未复合的抗体不干扰该测定。整个过程非常快速,适用于高通量筛选,而不需要从样品中生产蛋白质或纯化免疫球蛋白。使用这些方法,我们发现来自健康个体的血浆含有数百种与细胞蛋白质复合的自身抗体。因此,这种高度敏感的多重方法能够发现新的自身抗体-抗原或循环免疫复合物,其中许多可能用于疾病检测和表征。
We report on a novel, high-dimensional method to detect autoantibodies that are complexed with their natural autoantigens. Specifically, autoantibody-autoantigen complexes in serum or plasma are directly incubated onto a high-density antibody microarray. Detection of the bound autoantibody-antigen complex is made via fluorescently labeled anti-human immunoglobulin G or other immunoglobulin isotype secondary antibodies and quantification in a microarray scanner. Uncomplexed antibodies do not interfere with this assay. The whole process is very rapid and applicable for high-throughput screening without the need for production of proteins or immunoglobulin purification from the samples. Using these methods, we found that plasma from healthy individuals contains hundreds of autoantibodies complexed with cellular proteins. Thus, this highly sensitive, multiplex method is capable of discovering new autoantibody-antigen or circulating immune complexes many of which will likely be useful for disease detection and characterization.
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