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University of Nebraska - Lincoln

David Hage Publications

High-performance affinity chromatography

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Full-Text Articles in Physical Sciences and Mathematics

Development Of An On-Line Immunoextraction/Entrapment System For Protein Capture And Use In Drug Binding Studies By High-Performance Affinity Chromatography, Elliott L. Rodriguez, Saumen Poddar, Meera Choksi, David S. Hage Jan 2020

Development Of An On-Line Immunoextraction/Entrapment System For Protein Capture And Use In Drug Binding Studies By High-Performance Affinity Chromatography, Elliott L. Rodriguez, Saumen Poddar, Meera Choksi, David S. Hage

David Hage Publications

An on-line purification and entrapment system was developed that could extract a protein from a sample such as serum and entrap this protein within a small column for use in high-performance affinity chromatography. Human serum albumin (HSA) was employed as a model protein for this work. Immunoextraction columns containing polyclonal anti-HSA antibodies were developed to capture and isolate HSA from applied samples. This was followed by the use of a strong cation-exchange column to recapture and focus HSA as it eluted from the immunoextraction columns. The recaptured HSA was entrapped within 1.0 cm × 2.1 mm I.D. columns containing hydrazide-activated …


Affinity Chromatography: A Historical Perspective, David S. Hage, Ryan E. Matsuda Jan 2015

Affinity Chromatography: A Historical Perspective, David S. Hage, Ryan E. Matsuda

David Hage Publications

Affinity chromatography is one of the most selective and versatile forms of liquid chromatography for the separation or analysis of chemicals in complex mixtures. This method makes use of a biologically related agent as the stationary phase, which provides an affinity column with the ability to bind selectively and reversibly to a given target in a sample. This review examines the early work in this method and various developments that have lead to the current status of this technique. The general principles of affinity chromatography are briefly described as part of this discussion. Past and recent efforts in the generation …


Biointeraction Analysis By High-Performance Affinity Chromatography: Kinetic Studies Of Immobilized Antibodies, Mary Anne Nelson, Annette C. Moser, David S. Hage Jan 2010

Biointeraction Analysis By High-Performance Affinity Chromatography: Kinetic Studies Of Immobilized Antibodies, Mary Anne Nelson, Annette C. Moser, David S. Hage

David Hage Publications

A system based on high-performance affinity chromatography was developed for characterizing the binding, elution and regeneration kinetics of immobilized antibodies and immunoaffinity supports. This information was provided by using a combination of frontal analysis, split-peak analysis and peak decay analysis to determine the rate constants for antibody–antigen interactions under typical sample application and elution conditions. This technique was tested using immunoaffinity supports that contained monoclonal antibodies for 2,4-dichlorophenoxyacetic acid (2,4-D). Association equilibrium constants measured by frontal analysis for 2,4-D and related compounds with the immobilized antibodies were 1.7–12 × 106 M−1 at pH 7.0 and 25 °C. Splitpeak …