title: ProteoChip: a highly sensitive protein microarray prepared by a novel method of protein immobilization for application of protein-protein interaction studies.
abstract: We have developed a highly sensitive microarray protein chip, ProteoChip, coated with ProLinker, 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 B and that it is useful for studies of Ab-Ag interactions. ProteoChip, aminated glass slide coated with ProLinker 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.
authors.affiliation: Proteogen, RM 1516 Windstone, Yangjae-dong, Seocho-ku, Seoul, Korea.
authors.forename: Yoonsuk
authors.initials: Y
authors.lastname: Lee
authors.forename: Eun Kyoung
authors.initials: EK
authors.lastname: Lee
authors.forename: Yong Wan
authors.initials: YW
authors.lastname: Cho
authors.forename: Takuya
authors.initials: T
authors.lastname: Matsui
authors.forename: In-Cheol
authors.initials: IC
authors.lastname: Kang
authors.forename: Tai-Sun
authors.initials: TS
authors.lastname: Kim
authors.forename: Moon Hi
authors.initials: MH
authors.lastname: Han
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques / Q000379:methods*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D040081:Protein Array Analysis; D011485:Protein Binding; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D003432:Cross-Linking Reagents; D006046:Gold