期刊
PROTEOMICS
卷 3, 期 12, 页码 2289-2304出版社
WILEY
DOI: 10.1002/pmic.200300541
关键词
ProLinker; protein immobilization; protein microarray; protein-protein interaction; ProteoChip
We have developed a highly sensitive microarray protein chip, ProteoChip, coated with ProLinker(TM), novel calixcrown derivatives with a bifunctional coupling property that permits efficient immobilization of capture proteins on solid matrixes and makes high-throughput analysis of protein-protein interactions possible. The analysis of quartz crystal microbalance showed that both monoclonal antibody (mAb) and antigen (Ag) bound to the gold film of the sensor surface coated with ProLinker(TM) B and that it is useful for studies of Ab-Ag interactions. ProteoChip, aminated glass slide coated with ProLinker(TM) A, was also demonstrated to be useful for preparation of high-density array spots by using a microarrayer and for analysis of analyte Ags either by direct or sandwich methods of fluorescence immunoassay. The detection sensitivity of ProteoChip was as low as 1-10 femtogram/mL of analyte protein, useful for detection of tumor markers. ProteoChip was also useful for studies of direct protein-protein interactions as demonstrated by analysis of integrin-extracellular matrix protein interaction. These experimental results suggest that ProteoChip is a powerful tool for development of chip-based lead screening microarrays to monitor protein-protein interactions (i.e. drug target) as well as for biomarker assays which require high detection sensitivity.
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