Photoaffinity Immobilization of Fusion Proteins

This proposal presents an approach to resolving a critical and enduring obstacle to developing biosfor the timely detection of toxic and infectious agents. We will develop a passivated sensor surfaceuniquely with the specific binding protein in its crude source solution and, upon activation by lighprotein functionally oriented in a stabilizing, biomolecule-compatible environment. A trifunctionalderivatized on one site with a photoactivatable reagent capable upon illumination of forming covalenany organic target group (including all proteins) under mild reaction conditions. This reagent willbonded to a ligand for the specific binding protein and to the sensor surface previously passivatedphotoreactive hydrogel, to provide an essentially aqueous environment containing chosen ligand and padjacent on the sensor surface. These reagents are especially useful with antibodies and fusion protengineering technology. This approach thus presents an innovative combination of affinity chromatogrphotoaffinity labeling technologies to provide a cost-effective link between genetic engineering pro