The main objective of this project is to develop membranes with surfaces tailored for specific affinity for a particular protein or nucleic acid molecule. These 'smart' membranes will be used in the next generation of high throughput Layered Expression Scanning (LES) applications, such as tissue screening and analytical blotting (northern and western). The heart of this technology is a stack of novel membranes that allows a sample to be transferred directly through the stack creating nearly identical replicas. The membranes must be moderately porous and permeable, must allow for less than 5% lateral diffusion, must have high specificity of binding, must have high affinity for binding, and should be easily fabricated. During Phase I of this proposal, we will (1) construct 'smart' membrane prototypes (specific for 5 nucleic acid sequences) and will demonstrate their ability to specifically, and in a concentration-dependent manner, capture individual nucleic acid (NA) sequences from (i) liquid samples and (ii) gel samples following separation by electrophoresis, and (2) optimize a 'smart' membrane stack for capturing targeted nucleic acids. In Phase II, we plan to extend the basic platform concept to selective protein binding using antibodies tethered to the membrane surface through specific biological interactions and develop a diagnostic platform that can be used for molecular profiling of actual clinical samples. ? ?
