Photoaffinity Immobilization of Fusion Proteins

该提案提出了一种解决开发可靠的生物传感器系统以及时检测有毒和传染性病原体的关键和持久障碍的方法。这一装置对于在工业和人口稠密的环境中保护军事人员和平民至关重要。BSI公司提出开发一种钝化的传感器表面，该表面将与其粗源溶液中的特异性结合蛋白唯一地缔合，并且在被光活化时，共价键合该蛋白质，该蛋白质在功能上取向于稳定，生物分子-三官能氨基酸将在一个位点上用能够在照射时与基本上任何有机靶基团形成共价键的光活化试剂衍生化（包括所有蛋白质）在温和的反应条件下进行。然后，该试剂将热化学键合到特异性结合蛋白的配体上，并键合到先前用BSI的Photolink光反应性水凝胶钝化的传感器表面上，以提供基本上含水的环境，该环境含有选定的配体和传感器表面上相邻的光反应性基团。这些试剂特别适用于来自基因工程技术的抗体和融合蛋白。因此，这种方法提出了亲和层析和光亲和标记技术的创新组合，以提供基因工程产品和传感器设备之间的成本效益的链接。

This proposal presents an approach to resolving a critical and enduring obstacle to developing dependable biosensor systems for the timely detection of toxic and infectious agents. This device is vital to the protection of military personnel and of civilians in industrial and densely populated environments. BSI Corporation proposes to develop a passivated sensor surface which will associate uniquely with the specific binding protein in its crude source solution and, upon activation by light, covalently bond this protein functionally oriented in a stabilizing, biomolecule-compatible environment A trifunctional amino acid will be derivatized on one site with a photoactivatable reagent capable upon illumination of forming covalent bonds with essentially any organic target group (including all proteins) under mild reaction conditions. This reagent will then be thermochemically bonded to a ligand for the specific binding protein and to the sensor surface previously passivated with BSI's Photolink photoreactive hydrogel, to provide an essentially aqueous environment containing chosen ligand and photoreactive groups adjacent on the sensor surface. These reagents are especially useful with antibodies and fusion proteins from genetic engineering technology. This approach thus presents an innovative combination of affinity chromatography and photoaffinity labeling technologies to provide a cost-effective link between genetic engineering products and sensor devices.