Analysis of protein interactions on protein arrays by a wavelength interrogation-based surface plasmon resonance biosensor.
We have investigated whether surface plasmon resonance (SPR) sensors based on the wavelength interrogation are able to analyze protein interactions on protein arrays. The spectral SPR sensor was self-constructed and its detection limit, expressed as the minimal refractive index variation, was calculated to be 6.6x10(-5) with the signal fluctuation of 1.0x10(-5). The protein array surface was modified by a mixed thiol monolayer to immobilize proteins. Protein arrays were analyzed by the line-scanning mode of the SPR sensor, which scanned every 100 microm along the central line of array spots and the scanned results were presented by color spectra from blue to red. Glutathione S-transferase (GST)-rac1 caused a concentration-dependent increase of SPR wavelength shift on protein arrays. The surface structure of the protein arrays was analyzed by atomic force microscopy. Specific interactions of antigens with antibodies were analyzed on the protein arrays by using three antibodies and eight proteins. These results suggest that the wavelength interrogation-based SPR sensor can be used as the biosensor for the high-throughput analysis of protein interactions on protein arrays.
Yuk
Jong Seol
JS
Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine, Chunchon, Kangwon-Do, South Korea.
Jung
Se-Hui
SH
Jung
Jae-Wan
JW
Hong
Duk-Geun
DG
Han
Jeong-A
JA
Kim
Young-Myeong
YM
Ha
Kwon-Soo
KS
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism; D018625:Microscopy, Atomic Force; D040081:Protein Array Analysis; D025941:Protein Interaction Mapping; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013997:Time Factors; D020830:rac1 GTP-Binding Protein / Q000737:chemistry / Q000378:metabolism
D000906:Antibodies; D000941:Antigens; D011506:Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D020830:rac1 GTP-Binding Protein