期刊
PROTEOMICS
卷 6, 期 10, 页码 3199-3209出版社
WILEY
DOI: 10.1002/pmic.200500673
关键词
antibody microarrays; autoantibodies; autoantigens; biomarker; prostate cancer
资金
- NIDDK NIH HHS [U01 DK 063665] Funding Source: Medline
Diagnosing cancers based on serum profiling is a particularly attractive concept. However, the technical challenges to analysis of the serum proteome arise from the dynamic range of protein amounts. Cancer sera contain antibodies that react with a unique group of autologous cellular antigens, which affords a dramatic amplification of signal in the form of antibodies relative to the amount of the corresponding antigens. The serum autoantibody repertoire from cancer patients might, therefore, be exploited for antigen-antibody profiling. To date, studies of antigen-antibody reactivity using microarrays have relied on recombinant proteins or synthetic peptides as arrayed features. However, recombinant proteins and/or synthetic peptides may fail to accurately detect autoantibody binding due to the lack of proper PTMs. Here we describe the development and use of a reverse capture autoantibody microarray. Our reverse capture autoantibody microarray is based on the dual-antibody sandwich immunoassay platform of ELISA, which allows the antigens to be immobilized in their native configuration. As proof-of-principle, we demonstrate its use for antigen-autoantibody profiling with sera from patients with prostate cancer and benign prostate hyperplasia.
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