Analysis of protein interactions on protein arrays by a novel spectral surface plasmon resonance imaging.
We presented a novel surface plasmon resonance (SPR) imaging method for analysis of protein arrays based on a wavelength interrogation-based SPR biosensor. The spectral imaging was performed by the combination of position control and resonance wavelengths calculated from SPR reflectivity spectra. The imaging method was evaluated by analyzing interactions of glutathione S-transferase-fusion proteins with their antibodies. Antigen-antibody interactions were successfully analyzed on glutathione S-transferase-fusion protein arrays by using the spectral imaging method, and the results were confirmed by a parallel analysis using a previously used spectral SPR biosensor based on wavelength interrogation. Specific binding of anti-Rac1 and anti-RhoA to Rac1 and RhoA on the protein arrays was qualitatively and quantitatively analyzed by the spectral SPR imaging. Thus, it was suggested that the novel spectral SPR imaging was a useful tool for the high-throughput analysis of protein-protein interactions on protein arrays.
Yuk
Jong Seol
JS
Department of Molecular and Cellular Biochemistry and Nano-Bio Sensor Research Center, Kangwon National University College of Medicine, Chunchon, South Korea.
Kim
Hyun-Soo
HS
Jung
Jae-Wan
JW
Jung
Se-Hui
SH
Lee
Seung-Joon
SJ
Kim
Woo Jin
WJ
Han
Jeong-A
JA
Kim
Young-Myeong
YM
Ha
Kwon-Soo
KS
D000936:Antigen-Antibody Complex / Q000032:analysis*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D025941:Protein Interaction Mapping / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods
D000936:Antigen-Antibody Complex