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A Rational Approach for the Design and Synthesis of Selective Displacers for the Chromatographic Purification of Proteins

Srinavya Vutukuru1, Sandesh D. Kate1, Scott A. McCallum2, Christopher J. Morrison1, Steven M. Cramer1, and Ravi S. Kane1. (1) Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, (2) Department of Biology, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180

The development of effective strategies for designing selective displacers is critical for the successful implementation of displacement chromatography a powerful technique for the purification of biomolecules. We have developed an affinity-based approach for designing selective displacers, based on the identification of a protein-binding moiety and its attachment to a resin-binding moiety via a linker. In particular, we synthesized displacers based on biotin, which selectively retained avidin as compared to aprotinin on SP Sepharose High Performance resin. In addition, we have extended this approach to develop an affinity-peptide-based displacer that discriminates between lysozyme and cytochrome c. Our approach may be broadly applicable for the design of selective displacers for many proteins of biological or pharmaceutical interest. Combinatorial methods such as phage display are well-established and enable the rapid identification of affinity ligands; the attachment of these ligands to resin-binding moieties allows the immediate design of selective displacers.