This project deals with the development of a new type of stationary phase for chromatography and simulating moving bed (SMB) technology based on cross-linked protein crystals. While existing stationary phases can separate a wide variety of compounds, including mixtures of racemates, there are several limitations such as low loading, eluent limitations, narrow operational conditions and high cost, that preclude wider applications of chromatography. It is believed that some of these limitations can be successfully addressed by using Cross-Linked Enzyme Crystals (CLECá) or more broadly Cross-Linked Protein Crystals (CLPC). We have already demonstrated that cross-linked protein crystals are very stable, have porous structure, and demonstrate great affinity and chiral selectivity. In addition, they are mechanically stable and can be produced in large quantities. These crystals may have excellent performance characteristics as a new porous material and provide unique opportunities in at least three main types of liquid chromatography: size exclusion, affinity and chiral chromatography. In the Phase I study, the feasibility of using the five existing CLEC catalysts of lipases from Candida rugosa and Pseudomonas cepacia, subtilisin, thermolysin and penicillin acylase and a new CLPC derived from serum albumins as stationary phases for liquid chromatography will be demonstrated. The portion of synthetic chiral pharmaceuticals introduced as single enantiomers represents about $15 billion in sales and is expected to reach $150 billion by the end of the century. Chiral stationary phases could play and important role in producing and analyzing many of the final chemicals or intermediate used as optically pure drugs. The high price of bulk CSP currently precludes the wide use of this technology in either preparative chromatography or simulating moving bed (SMB) technology. This situation may dramatically change with the introduction of CSP based on protein crystals.
