Columns, Media, and Methods for Ultrafast Affinity Chromatography

The US liquid chromatography market earned $596.3 million in 2006, 44.4% of which came from consumables such as columns and resins. Technological innovation is predicted to drive sales of these consumables to $473.2 million by 2013 (Source).

Dr. David Hage and colleagues at the University of Nebraska–Lincoln have developed ultra-rapid high-performance affinity chromatography (HPAC) and affinity capillary electrophoresis (ACE) methods for biological and environmental analysis. These methods use novel microcolumns to separate samples with times ranging from less than a second to several minutes. New classes of chromatographic media offer improved selectivity and biological relevance.

• Sub-millimeter length columns can separate analytes within milliseconds under standard HPLC conditions
• Novel chromatographic media combine the benefits of affinity and size-exclusion chromatography to distinguish between free and protein-bound analytes
• New immobilization strategies allow biological ligands to mimic in vivo behavior.

• Clinical measurement of free (i.e., biologically active) drug concentration in blood or serum for individualized dosage determination
• Measurement of environmental contaminants in water
• High-throughput screening of combinatorial chemistry libraries and drug candidates
• Protein-based chiral separations
• Study of protein-drug interactions to predict in vivo drug behavior

• Clarke, W., Hage, D.S. (2001) "Development of Sandwich HPLC Microcolumns for Analyte Adsorption on the Millisecond Time Scale" Anal Chem. 73:1366-1373.
• Clarke, W., Choudhuri, A.R., Hage, D.S. (2001) "Analysis of Free Drug Fractions by Ultra-fast Immunoaffinity Chromatography" Anal. Chem.73:2157-2164.
• Wa, C., Mallik, R., Hage, D.S. (2008) "Development of Immunoaffinity Restricted Access Media for Rapid Extractions of Low-Mass Analytes" Anal. Chem.80:8751-8762.
• Hage, D.S. et. al. (2009) "Characterization of drug–protein interactions in blood using high–performance affinity chromatography" J. Sep. Sci. 32:835-853.