Introduction of Capillary Electrophoresis
The introduction of capillary zone electrophoresis is a major advance in serum protein electrophoresis. Referred as high-performance capillary electrophoresis (HPCE), capillary zone electrophoresis (CZE), Free solution capillary electrophoresis (FSCE), and Capillary electrophoresis (CE).
Instrumentation
Instrument includes-Capillary outlet
Capillary inlet
Capillary tube
High voltage supply
Anode
cathode
Reservoir
Electrolyte buffer
Detector
Data acquisition as shown above image
Procedure of Capillary electrophoresis
- Fill the tube with the sample solution.
- Carry out the sample application by high voltage injection or by pressure injection.
- Apply a high voltage of 20kV.
- Two forces act on the proteins: The attraction of first the negatively charged proteins toward the positive electrode anode and second, an electro-osmotic force (EOF).
- EOF drags all the molecules towards the cathode irrespective of the charge.
- During this migration, positively charged molecules move faster than negatively charged ones. Detection of resolved molecules is done by a UV monitor and the signal is recording in a recorder.
- A capillary electrophoretogram can obtain by plotting the signal against time.
Advantages
- Automation
- High speed
- High voltage can be applied
- Proper heat dissipation
- A small amount of sample is enough
Applications of Capillary electrophoresis
- Used to separate:
- Inorganic ions
- Amino acids
- Organic acids
- Drug
- Vitamins
- Porphyrins
- Carbohydrates
- Oligonucleotide
- Proteins
- DNA
Further Readings
- Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 5th Edition
- Principles and Techniques of Biochemistry and Molecular Biology, Wilson and Walker 7th Edition
- Short protocols in molecular biology, 4th edition
- Pulsed-field gel electrophoresis( A practical Guide )by Bruce Birren21.
- Molecular Biology. Editor: David Freifelder, 2nd ed, Publisher Jones and Bartlett Inc.