IMMOBILIZATION CHEMISTRIES SUITABLE FOR USE IN THE BIACORE SURFACE-PLASMON RESONANCE DETECTOR
IMMOBILIZATION CHEMISTRIES SUITABLE FOR USE IN THE BIACORE SURFACE-PLASMON RESONANCE DETECTOR
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DOI：
10.1016/0003-2697(92)90589-y
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发表时间：
1992-08-15
影响因子：
2.9
通讯作者：
PECK, K
中科院分区：
文献类型：
--
作者：
OSHANNESSY, DJ;BRIGHAMBURKE, M;PECK, K
表面等离子体共振检测器，如由Pharmacia生产的BIAcore仪器，显示出以无标记模式检测和定量大分子相互作用的前景。这种检测器依赖于相互作用物质之一共价固定到感测表面上。迄今为止，唯一公开的用于此目的的化学是含伯氨基的配体与N-羟基琥珀酰亚胺（NHS）酯活化的表面的反应。为了增加BIAcore在固定配体方面的多功能性，我们对与该系统相容的活化化学进行了研究。使用容易获得的试剂，我们证明了羧化葡聚糖包被的传感表面可以很容易地转化为NHS酯以外的功能，包括胺活化，肼活化和巯基活化的表面。此外，利用链霉亲和素/生物素相互作用，通过采用特异性修饰的生物素衍生物来探测感测表面的化学修饰。
Surface plasmon resonance detectors, such as the BIAcore instrument produced by Pharmacia, show promise for the detection and quantitation of macromolecular interactions in a label-free mode. Such detectors rely on the covalent immobilization of one of the interacting species onto the sensing surface. To date, the only published chemistry for this purpose is reaction of primary amino-containing ligands with an N-hydroxysuccinimide (NHS) ester-activated surface. In an effort to increase the versatility of the BIAcore with respect to immobilizing ligands, we undertook an investigation of activation chemistries compatible with this system. Using readily available reagents, we demonstrated that the carboxylated dextran-coated sensing surface could be easily converted to functions other than NHS-esters, including amine-activated, hydrazine-activated, and sulfhydryl-activated surfaces. In addition, use was made of the streptavidin/biotin interaction to probe chemical modifications of the sensing surface, by employing specifically modified biotin derivatives.