Micro-Scale Frontal Affinity Chromatography with Mass Spectrometric Detection: A New Method for the Screening of
David C Schriemer1, David R Bundle1, Liang Li1
1Department of Chemistry, University of Alberta, Edmonton, T6G 2G2 (Canada), Fax: (+1) 403-492-7705.
Angewandte Chemie (International Ed. in English)
|May 2, 2018
Summary
This study introduces a method to determine binding constants (Kd) for multiple ligands simultaneously using affinity chromatography and mass spectrometry. This technique efficiently quanties ligand-receptor interactions with minimal sample amounts.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Pharmacology
Background:
- Quantifying ligand-receptor binding is crucial for drug discovery and understanding biological processes.
- Traditional methods for determining binding constants can be time-consuming and require significant amounts of purified compounds.
Purpose of the Study:
- To develop a high-throughput method for determining binding constants (Kd) of multiple ligands.
- To enable the analysis of small sample quantities (sub-microgram).
Main Methods:
- Utilizing affinity chromatography with an immobilized receptor.
- Infusing a mixture of ligands continuously through the column.
- Analyzing the eluent using electrospray mass spectrometry.
- Generating an affinity chromatogram to determine breakthrough volumes.
Main Results:
- Successfully determined binding constants (Kd) for all active components in a ligand library in a single run.
- Demonstrated the ability to quantify individual compound affinities from a mixture.
- Achieved determination with sub-microgram quantities of ligands.
Conclusions:
- The developed method offers an efficient approach for characterizing ligand-receptor interactions.
- This technique facilitates rapid screening of ligand libraries.
- It is suitable for applications requiring minimal sample input and high throughput.
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