ProteoChip: a highly sensitive protein microarray prepared by a novel method of protein immobilization for application of protein-protein interaction studies.
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.
Lee
Yoonsuk
Y
Proteogen, RM 1516 Windstone, Yangjae-dong, Seocho-ku, Seoul, Korea.
Lee
Eun Kyoung
EK
Cho
Yong Wan
YW
Matsui
Takuya
T
Kang
In-Cheol
IC
Kim
Tai-Sun
TS
Han
Moon Hi
MH
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
D000911:Antibodies, Monoclonal; D000941:Antigens; D003432:Cross-Linking Reagents; D006046:Gold