Basic Information
Abstract Number: 2130-1    
Author Name: David S Hage Affiliation: University of Nebraska
Session Title: Affinity Methods in Biochemical Separations
Event Type: Symposia
Event Title: Recent Advances and Applications of Microaffinity Chromatography
Presider(s): Woolley, Adam T Start Time: 02:05 PM ( Slot # 2 )
Date: Wednesday, March 3rd, 2010 Location: 308B
Keywords: Bioanalytical, Clinical Chemistry, HPLC, Separation Sciences

Abstract Content
Affinity ligands have been used for decades for both the selective isolation and analysis of sample components in complex mixtures. The selectivity and strong binding of many biological agents that are used as affinity ligands, such as antibodies, have also made these ligands of great interest over the last decade for use in HPLC methods and, more recently, in microaffinity LC methods. This presentation will examine some recent developments in the creation and bioanalytical applications of affinity microcolumns. One set of methods that will be discussed are chromatographic-based immunoassay, in which antibodies or related agents have been used in a variety of detection formats for the rapid analysis or drugs, hormones, proteins, and other analytes. Affinity microcolumns are particularly valuable in the area of ultrafast immunoextraction, in which analytes are isolated from samples in the sub-second time domain. One unique application that will be discussed for this approach is in the analysis of free drug and drug and hormone fractions in blood. Several approaches for conducting these assays will also be examined, including a number of competitive or displacement assay formats and the use of either packed microcolumns or small affinity monoliths. Affinity microcolumns have been shown to be valuable in the studying the thermodynamics and kinetics of biological interactions. This type of application will be illustrated by using examples based on the binding of drugs with serum agents such as human serum albumin HSA, alpha 1-acid glycoprotein and lipoproteins. The benefits and limitations of using affinity microcolumns for this purpose will be examined and the use of this approach in the high-throughput screening of biological interactions will be discussed. This work was funded by the NIH under grant R01 GM 044931.