In the first grant we demonstrated that novel low molecular weight (MW) displacers can be successfully employed for high resolution/high throughput protein purifications in ion exchange systems. We have recently demonstrated that low molecular weight displacers can also be used for oligonucleotide purification and that the stationary phase has a significant effect on the efficacy of various low molecular weight displacers. In this phase of the project we will expand this research by addressing several critical issues related to the rational design of efficient low MW displacers. The proposed research will expand our investigation into the relationship between displacer chemistry and its efficacy as a protein displacer. The proposed research will include displacer synthesis, development of high affinity displacers for cation and anion exchange systems, development of efficient selective displacers, and the application of these variants of a variety of low MW displacers identified in the first grant. In addition, this phase of the project will examine the effect of stationary phase chemistry on the efficacy of various classes of low MW displacers using resins with various backbone chemistry, spacer arm chemistry, charged ligands and ligand densities. The efficacy of displacement chromatography for oligonucleotide purifications will be examined in detail. The displacers developed in this research will be employed in a detailed investigation of the interplay between displacer chemistry and displacer affinity on various stationary phase materials. This research will evaluate several hypotheses related to displacer design. The low MW displacers developed in this research will be employed for the purification of proteins and oligonucleotides from complex biological mixtures. Finally, these high affinity displacers will be employed to develop a generic displacement will provide significant insight into the design or efficient low MW displacers for protein and oligonucleotide purification and will enable the purification of a wide range of biopharmaceuticals in a more efficient manner. Furthermore, this work will facilitate the use of this high resolution/high throughput technique by medical researchers at the bench scale for the rapid purification of novel biomolecules form dilute complex mixtures.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Special Emphasis Panel (ZRG1-BMT (01))
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Edmonds, Charles G
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Rensselaer Polytechnic Institute
Engineering (All Types)
Schools of Engineering
United States
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Levy, Miriam H; Plawsky, Joel; Cramer, Steven M (2013) Photopolymerized sol-gel monoliths for separations of glycosylated proteins and peptides in microfluidic chips. J Sep Sci 36:2358-65
Evans, Steven T; Holstein, Melissa; Cramer, Steven M (2011) Detection of trace proteins in multicomponent mixtures using displacement chromatography. Anal Chem 83:4184-92
Evans, Steven T; Morrison, Christopher J; Freed, Alexander et al. (2010) The effect of feed composition on the behavior of chemically selective displacement systems. J Chromatogr A 1217:1249-54
Morrison, Christopher J; Godawat, Rahul; McCallum, Scott A et al. (2009) Mechanistic studies of displacer-protein binding in chemically selective displacement systems using NMR and MD simulations. Biotechnol Bioeng 102:1428-37
Evans, Steven T; Freed, Alexander; Cramer, Steven M (2009) Displacer concentration effects in displacement chromatography. Implications for trace solute detection. J Chromatogr A 1216:79-85
Morrison, Christopher J; Cramer, Steven M (2009) Characterization and design of chemically selective cationic displacers using a robotic high-throughput screen. Biotechnol Prog 25:825-33
Morrison, Christopher J; Park, Sun Kyu; Simocko, Chester et al. (2008) Synthesis and characterization of fluorescent displacers for online monitoring of displacement chromatography. J Am Chem Soc 130:17029-37
Liu, Jia; Hilton, Zachary A; Cramer, Steven M (2008) Chemically selective displacers for high-resolution protein separations in ion-exchange systems: effect of displacer-protein interactions. Anal Chem 80:3357-64
Rege, Kaushal; Viswanathan, Gunaranjan; Zhu, Guangyu et al. (2006) In vitro transcription and protein translation from carbon nanotube-DNA assemblies. Small 2:718-22
Rege, Kaushal; Ladiwala, Asif; Hu, Shanghui et al. (2005) Investigation of DNA-binding properties of an aminoglycoside-polyamine library using quantitative structure-activity relationship (QSAR) models. J Chem Inf Model 45:1854-63

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