PROJECT SUMMARY The development of methods for the separation, analysis and resolution of chiral drugs is of important interest for pharmaceutical development. Two enantiomers of the same compound may have the same physical or chemical properties, but show marked differences in how they interact in a human system (i.e., regarding their pharmacology, toxicology, pharmacokinetics and metabolism). Thus, the chiral separation of drugs is important to eliminate the unwanted enantiomer from the preparation. Separation of enantiomers is typically achieved by high- performance liquid chromatography (HPLC) where a chiral selector is used in a chiral stationary phase (CSP). CSPs can be composed of several types of molecules, including polysaccharides (i.e., amylose and cellulose) and proteins (e.g., albumin, enzymes), among others. Only a handful of proteins have been investigated for use as HPLC CSPs despite their unique enantioselective properties. Unfortunately current, protein-based CSPs have low loading capacity, are expensive to prepare, and have limited stability. There is clearly a market need for enzyme and protein- based CSP separation methods with higher loading capacity, less degradation and extended shelf life. Based on market research interviews that we have conducted through NSF’s I-Corps program on our platform biomaterial technology, it was made clear that the technology for chiral columns has not changed and that innovation in the field was desired. Bondwell Technologies aims to develop a novel high-capacity protein-based selector for use as a CSP based on our innovative biomaterial. The unique qualities of our biomaterial addresses all of current limitations of chiral CBHs. During this proposed Phase I feasibility effort, we will first express the chiral selector enzyme cellobiohydrolase (CBH I) and form our biomaterials, We will test the enzyme in the biomaterial for function to ensure correct folding. We will then prepare our material to be used as a CSP. Finally, we will test for the ability of our biomaterial to separate enantiomers of the beta- bocking drugs, propranolol and alprenolol.