On-chip capillary electrophoresis for alkaline phosphatase testing.

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Biochemistry

Background:

• Capillary electrophoresis (CE) is a powerful separation technique.
• On-chip microfluidic devices offer advantages for rapid analysis.
• Detecting specific enzyme activity in complex biological samples remains challenging.

Purpose of the Study:

• To develop an integrated on-chip capillary electrophoresis system for blood analysis.
• To detect alkaline phosphatase (ALP) activity in a mixture of proteins.
• To utilize laser-induced fluorescence for sensitive enzyme detection.

Main Methods:

• Photolithographic fabrication of a capillary electrophoresis device on a glass chip.
• Electrophoretically mediated microanalysis (EMMA) for enzyme detection.
• Use of fluorescein diphosphate as a fluorogenic substrate for ALP.
• Laser-induced fluorescence monitoring of fluorescein production.

Main Results:

• Successful fabrication of an on-chip capillary electrophoresis device.
• Detection of small amounts of alkaline phosphatase (ALP) enzyme.
• Demonstration of EMMA for analyzing enzyme activity in protein mixtures.
• Optimization of methods to minimize protein adhesion on the chip.

Conclusions:

• The developed on-chip CE device is suitable for sensitive blood analysis.
• The EMMA method combined with fluorescence detection enables specific enzyme activity measurement.
• This technology has potential for point-of-care diagnostics and biochemical assays.