Analysis of protein interactions on protein arrays by a wavelength interrogation‐based surface plasmon resonance biosensor
Analysis of protein interactions on protein arrays by a wavelength interrogation‐based surface plasmon resonance biosensor
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DOI：
10.1002/pmic.200400879
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发表时间：
2004-11
期刊：
影响因子：
3.4
通讯作者：
J. Yuk;Se-Hui Jung;Jae-Wan Jung;D. Hong;J. Han;Young-Myeong Kim;K. Ha
中科院分区：
文献类型：
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作者：
J. Yuk;Se-Hui Jung;Jae-Wan Jung;D. Hong;J. Han;Young-Myeong Kim;K. Ha
我们研究了基于波长探询的表面等离子体共振（SPR）传感器是否能够在蛋白质阵列上分析蛋白质相互作用。光谱SPR传感器是自行构建的，其检测限（表示为最小折射率变化）计算为6.6×10−5，信号波动为1.0×10− 5。蛋白质阵列表面通过混合硫醇单层修饰以包裹蛋白质。利用SPR传感器的线扫描模式对蛋白质阵列进行分析，沿阵列点的中心线沿着每100 μm扫描一次，扫描结果由蓝色到红色的彩色光谱呈现。谷氨酰胺S-转移酶（GST）-rac 1引起蛋白质阵列上SPR波长偏移的浓度依赖性增加。用原子力显微镜分析了蛋白质阵列的表面结构。通过使用三种抗体和八种蛋白质在蛋白质阵列上分析抗原与抗体的特异性相互作用。这些结果表明，基于波长询问的SPR传感器可以用作蛋白质阵列上蛋白质相互作用的高通量分析的生物传感器。
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 6.6×10−5 with the signal fluctuation of 1.0×10−5. 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 μm 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.