TY - JOUR
T1 - Analysis of biomolecular interactions using affinity microcolumns
T2 - A review
AU - Zheng, Xiwei
AU - Li, Zhao
AU - Beeram, Sandya
AU - Podariu, Maria
AU - Matsuda, Ryan
AU - Pfaunmiller, Erika L.
AU - White, Christopher J.
AU - Carter, Na Tasha
AU - Hage, David S.
N1 - Publisher Copyright:
© 2014 Elsevier B.V..
PY - 2014/9/15
Y1 - 2014/9/15
N2 - 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.
AB - 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.
KW - Affinity microcolumns
KW - Biointeraction analysis
KW - Frontal affinity chromatography
KW - High-performance affinity chromatography
KW - Ultrafast affinity extraction
KW - Zonal elution
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U2 - 10.1016/j.jchromb.2014.01.026
DO - 10.1016/j.jchromb.2014.01.026
M3 - Review article
C2 - 24572459
AN - SCOPUS:84927172776
SN - 1570-0232
VL - 968
SP - 49
EP - 63
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
ER -