Automated microanalysis using magnetic beads with commercial capillary electrophoretic instrumentation

• Published in: Journal of Chromatography A 1997-09, Vol.781 (1), p.197-204
• Main Authors: Rashkovetsky, Leonid G., Lyubarskaya, Yelena V., Foret, Frantisek, Hughes, Dallas E., Karger, Barry L.
• Format: Article
• Language: English
• Subjects: AIDS/HIV; Alkaline Phosphatase - analysis; Alkaline Phosphatase - chemistry; Alkaline Phosphatase - immunology; Analytical, structural and metabolic biochemistry; Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; Antigens; Biological and medical sciences; Electrophoresis, Capillary - methods; Enzymatic analysis; Enzymes; Enzymes and enzyme inhibitors; Enzymes, Immobilized - analysis; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - immunology; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; HIV Protease - analysis; HIV Protease - chemistry; HIV Protease Inhibitors - chemistry; Immunoassays; Magnetic beads; Magnetics; Mice; Microspheres; Osmolar Concentration; Pepstatins - chemistry; Spectrometry, Fluorescence; Time Factors
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(97)00629-8

The potential of a new microanalytical method using magnetic beads (MBs) and commercial capillary electrophoresis (CE) instrumentation for performing enzymatic and inhibition assays, as well as for analysis of biological molecules such as antigens, substrates, etc., has been explored. A small quantity of magnetic beads containing immobilized biomolecules was injected into a neutral hydrophilic-coated fused-silica capillary. The short plug (2–3 mm) of beads was held fixed by a magnet placed in the cartridge of the CE system, without the use of frits. The beads could be replaced after each run, eliminating the need to regenerate the solid support. Two protocols were used for analysis: sequential injection (SI) and SI followed by isotachophoretic (ITP) focusing. Alkaline phosphatase (AP) and HIV-protease were used to demonstrate the SI procedure for enzymatic and inhibition assays. The second protocol, SI/ITP, was employed to quantitate an antigen (mouse mAB) using antibodies (sheep IgG towards mouse AB) immobilized on the beads. The MB-CE method, requiring only femtomole (fmol) quantities of material, can potentially be employed in diagnostic and forensic assays, kinetic studies and searching for inhibitors, ligands, receptors, etc.