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Capillary Electrophoresis Separation of Monoclonal Antibody Isoforms Using a Neutral Capillary
Published on: January 16, 2017
Selective preconcentration for capillary zone electrophoresis using protein G immunoaffinity capillary chromatography
1Department of Chemistry, University of Florida, Gainesville 32611-7200, USA.
Electrophoresis
|April 1, 1995
Summary
This study introduces an immunoaffinity preconcentrator for capillary zone electrophoresis, enabling significant sample concentration. The method effectively isolates and enriches antigens from large volumes for improved analysis.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Immunoaffinity chromatography is crucial for selective analyte isolation.
- Capillary zone electrophoresis offers high-resolution separations.
- Integrating these techniques enhances analytical sensitivity.
Purpose of the Study:
- To develop an on-line and off-line immunoaffinity preconcentration system for capillary zone electrophoresis.
- To achieve significant preconcentration factors for analytes in complex matrices.
- To demonstrate the system's utility with insulin in serum samples.
Main Methods:
- Packing capillaries with protein G chromatographic support to create an immunoaffinity stationary phase.
- Utilizing selective retention and desorption of antigen-antibody complexes.
- Coupling the preconcentration column with capillary zone electrophoresis via a flow-gated interface (on-line) or fraction collection (off-line).
Main Results:
- Achieved 1000-fold preconcentration by loading 1 mL samples and desorbing in 1 microL.
- Demonstrated selective retention and preconcentration of antigen.
- Successfully applied the system to serum samples spiked with insulin.
Conclusions:
- The developed immunoaffinity preconcentrator significantly enhances sensitivity for capillary zone electrophoresis.
- The on-line and off-line configurations provide flexibility for various analytical workflows.
- This method is effective for analyzing low-abundance analytes like insulin in biological samples.
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