Photoaffinity Immobilization of Fusion Proteins

本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感染性病原体的关键和持久障碍的方法。这一装置对保护军事人员和工业环境和人口稠密环境中的平民至关重要。BSI公司建议开发一种钝化传感器表面，该表面将与其原始源溶液中的特定结合蛋白唯一地结合，并且在光激活后，将该蛋白以稳定的，在温和的反应条件下，三功能氨基酸将在一个位点上用光活化试剂衍生，该试剂在光照下能够与基本上任何有机靶基团（包括所有蛋白质）形成共价键。然后，该试剂将与特定结合蛋白的配体和先前用BSI的Photolink光反应性水凝胶钝化的传感器表面进行热化学结合，以提供一个基本的水环境，其中包含选定的配体和光反应性基团相邻的传感器表面。这些试剂对来自基因工程技术的抗体和融合蛋白特别有用。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，为基因工程产品和传感器设备之间提供了一种具有成本效益的联系。

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.