High-Throughput Screening for Native Autoantigen鈥揂utoantibody Complexes Using Antibody Microarrays
详细信息 查看全文
- 作者：Jung-hyun Rho ; Paul D. Lampe
- 刊名：Journal of Proteome Research
- 出版年：2013
- 出版时间：May 3, 2013
- 年：2013
- 卷：12
- 期：5
- 页码：2311-2320
- 全文大小：431K
- 年卷期：v.12,no.5(May 3, 2013)
- ISSN：1535-3907
文摘
We report on a novel, high-dimensional method to detect autoantibodies that are complexed with their natural autoantigens. Specifically, autoantibody鈥揳utoantigen complexes in serum or plasma are directly incubated onto a high-density antibody microarray. Detection of the bound autoantibody鈥揳ntigen complex is made via fluorescently labeled antihuman 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鈥揳ntigen or circulating immune complexes, many of which will likely be useful for disease detection and characterization.