Analysis of biomolecular interactions using affinity microcolumns: A review

作者：Zheng Xiwei; Li Zhao; Beeram Sandya; Podariu Maria; Matsuda Ryan; Pfaunmiller Erika L; White Christopher I II; Carter NaTasha; Hage David S^*

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2014, 968: 49-63.

DOI：10.1016/j.jchromb.2014.01.026

• Affinity microcolumns
• High-performance affinity chromatography
• Biointeraction analysis
• Zonal elution
• Frontal affinity chromatography
• Ultrafast affinity extraction

Affinity chromatography has become an important tool for characterizing biomolecular interactions. The use of affinity microcolumns, which contain immobilized binding agents and have volumes in the mid-to-low microliter range, has received particular attention in recent years. Potential advantages of affinity microcolumns include the many analysis and detection formats that can be used with these columns, as well as the need for only small amounts of supports and immobilized binding agents. This review examines how affinity microcolumns have been used to examine biomolecular interactions. Both capillary-based microcolumns and short microcolumns are considered. The use of affinity microcolumns with zonal elution and frontal analysis methods are discussed. The techniques of peak decay analysis, ultrafast affinity extraction, split-peak analysis, and band-broadening studies are also explored. The principles of these methods are examined and various applications are provided to illustrate the use of these methods with affinity microcolumns. It is shown how these techniques can be utilized to provide information on the binding strength and kinetics of an interaction, as well as on the number and types of binding sites. It is further demonstrated how information on competition or displacement effects can be obtained by these methods.

• 出版日期2014-10-1