Single Cells as Biosensors for Chemical Separations
Abstract
A biosensor system based on the response of living cells was demonstrated that can detect specific components of a complex mixture fractionated by a microcolumn separation technique. This system uses ligand-receptor binding and signal-transduction pathways to biochemically amplify the presence of an analyte after electrophoretic separation. The transduced signal was measured by means of two approaches: (i) fluorescence determination of intracellular calcium concentrations in one or more rat PC-12 cells and (ii) measurement of transmembrane current in a Xenopus laevis oocyte microinjected with messenger RNA that encodes a specific receptor. This analysis system has the potential to identify biologically active ligands present in a complex mixture with exceptional sensitivity and selectivity.
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References
ABDELLATIF, M.S., ENZYME-BASED FIBER OPTIC SENSOR FOR GLUCOSE DETERMINATION, ANALYTICAL LETTERS 21: 943 (1988).
BRIGGS, C.A., ACTIVATION OF THE 5-HT1C RECEPTOR EXPRESSED IN XENOPUS OOCYTES BY THE BENZAZEPINES SCH-23390 AND SKF-38393, BRITISH JOURNAL OF PHARMACOLOGY 104: 1038 (1991).
BUSH, A.B., NERVE GROWTH-FACTOR POTENTIATES BRADYKININ-INDUCED CALCIUM INFLUX AND RELEASE IN PC12 CELLS, JOURNAL OF NEUROCHEMISTRY 57: 562 (1991).
Cooper, J. R., The Biochemical Basis of Neuropharmacology 5 173 (1986).
DELORME, E.M., EFFECTS OF PROLONGED DEPOLARIZATION ON THE NICOTINIC ACETYLCHOLINE-RECEPTORS OF PC12 CELLS, JOURNAL OF NEUROCHEMISTRY 50: 1248 (1988).
FASOLATO, C, RECEPTOR-MEDIATED CALCIUM INFLUX IN PC12 CELLS - ATP AND BRADYKININ ACTIVATE 2 INDEPENDENT PATHWAYS, JOURNAL OF BIOLOGICAL CHEMISTRY 265: 20351 (1990).
FISHMAN, H.A., SPONTANEOUS INJECTION IN MICROCOLUMN SEPARATIONS, ANALYTICAL CHEMISTRY 66: 2318 (1994).
GANDHI, C.R., HEPATIC-EFFECTS OF ENDOTHELIN - RECEPTOR CHARACTERIZATION AND ENDOTHELIN-INDUCED SIGNAL TRANSDUCTION IN HEPATOCYTES, BIOCHEMICAL JOURNAL 287: 897 (1992).
GIAEVER, I, ANTIBODY-ANTIGEN REACTION - VISUAL OBSERVATION, JOURNAL OF IMMUNOLOGY 110: 1424 (1973).
GREANEY, M.D., IMPROVED METHOD FOR THE ROUTINE ANALYSIS OF ACETYLCHOLINE-RELEASE IN-VIVO - QUANTITATION IN THE PRESENCE AND ABSENCE OF ESTERASE INHIBITOR, JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS 622: 125 (1993).
Greene, L. A., Culturing Nerve Cells: 207 (1991).
GREENE, L.A., SYNTHESIS, STORAGE AND RELEASE OF ACETYLCHOLINE BY A NORADRENERGIC PHEOCHROMOCYTOMA CELL LINE, NATURE 268: 349 (1977).
JACKSON, T, STRUCTURE AND FUNCTION OF G-PROTEIN COUPLED RECEPTORS, PHARMACOLOGY & THERAPEUTICS 50: 425 (1991).
KIST, TBL, CAPILLARY ZONE ELECTROPHORESIS SEPARATION OF KININS USING A NOVEL LASER FLUORESCENCE DETECTOR, BRAZILIAN JOURNAL OF MEDICAL AND BIOLOGICAL RESEARCH 27: 11 (1994).
MONNIG, C.A., CAPILLARY ELECTROPHORESIS, ANALYTICAL CHEMISTRY 66: R280 (1994).
MORI, M, STRUCTURE OF CCF10, A PEPTIDE SEX-PHEROMONE WHICH INDUCES CONJUGATIVE TRANSFER OF THE STREPTOCOCCUS-FAECALIS TETRACYCLINE RESISTANCE PLASMID, PCF10, JOURNAL OF BIOLOGICAL CHEMISTRY 263: 14574 (1988).
NIE, S, unpublished data.
OATES, M.D., QUANTITATIVE AMINO-ACID-ANALYSIS OF INDIVIDUAL SNAIL NEURONS BY OPEN TUBULAR LIQUID-CHROMATOGRAPHY, ANALYTICAL CHEMISTRY 62: 1573 (1990).
OLEFIROWICZ, T.M., CAPILLARY ELECTROPHORESIS IN 2-MU-M AND 5-MU-M DIAMETER CAPILLARIES - APPLICATION TO CYTOPLASMIC ANALYSIS, ANALYTICAL CHEMISTRY 62: 1872 (1990).
PARCE, J.W., DETECTION OF CELL-AFFECTING AGENTS WITH A SILICON BIOSENSOR, SCIENCE 246: 243 (1989).
POZZAN, T, ACTIVATION OF MUSCARINIC RECEPTORS IN PC12 CELLS STIMULATION OF CA-2+ INFLUX AND REDISTRIBUTION, BIOCHEMICAL JOURNAL 234: 547 (1986).
ROGERS, K.R., ACETYLCHOLINE-RECEPTOR FIBER-OPTIC EVANESCENT FLUOROSENSOR, ANALYTICAL BIOCHEMISTRY 182: 353 (1989).
ROUNDS, M.A., ANION-EXCHANGE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH POSTCOLUMN DETECTION FOR THE ANALYSIS OF PHYTIC ACID AND OTHER INOSITOL PHOSPHATES, JOURNAL OF CHROMATOGRAPHY A 653: 148 (1993).
SANDS, S.B., CALCIUM PERMEABILITY OF NEURONAL NICOTINIC ACETYLCHOLINE-RECEPTOR CHANNELS IN PC12 CELLS, BRAIN RESEARCH 560: 38 (1991).
SMITH, R. D., ANALYTICAL CHEMISTRY 65: A574 (1993).
SUGAO, N, NA+/D-GLUCOSE COTRANSPORTER BASED BILAYER LIPID-MEMBRANE SENSOR FOR D-GLUCOSE, ANALYTICAL CHEMISTRY 65: 363 (1993).
SUZUKI, H, AN AMPEROMETRIC SENSOR FOR CARBON-DIOXIDE BASED ON IMMOBILIZED BACTERIA UTILIZING CARBON-DIOXIDE, ANALYTICA CHIMICA ACTA 199: 85 (1987).
Turner, A. P. F., Advances in Biosensors (1991).
UPDIKE, S.J., ENZYME ELECTRODE, NATURE 214: 986 (1967).
YAMAMOTO, K, J CHROMATOGR 588: 327 (1991).
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Science
Volume 267 | Issue 5194
6 January 1995
6 January 1995
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© 1995.
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Published in print: 6 January 1995
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References
References
References
ABDELLATIF, M.S., ENZYME-BASED FIBER OPTIC SENSOR FOR GLUCOSE DETERMINATION, ANALYTICAL LETTERS 21: 943 (1988).
BRIGGS, C.A., ACTIVATION OF THE 5-HT1C RECEPTOR EXPRESSED IN XENOPUS OOCYTES BY THE BENZAZEPINES SCH-23390 AND SKF-38393, BRITISH JOURNAL OF PHARMACOLOGY 104: 1038 (1991).
BUSH, A.B., NERVE GROWTH-FACTOR POTENTIATES BRADYKININ-INDUCED CALCIUM INFLUX AND RELEASE IN PC12 CELLS, JOURNAL OF NEUROCHEMISTRY 57: 562 (1991).
Cooper, J. R., The Biochemical Basis of Neuropharmacology 5 173 (1986).
DELORME, E.M., EFFECTS OF PROLONGED DEPOLARIZATION ON THE NICOTINIC ACETYLCHOLINE-RECEPTORS OF PC12 CELLS, JOURNAL OF NEUROCHEMISTRY 50: 1248 (1988).
FASOLATO, C, RECEPTOR-MEDIATED CALCIUM INFLUX IN PC12 CELLS - ATP AND BRADYKININ ACTIVATE 2 INDEPENDENT PATHWAYS, JOURNAL OF BIOLOGICAL CHEMISTRY 265: 20351 (1990).
FISHMAN, H.A., SPONTANEOUS INJECTION IN MICROCOLUMN SEPARATIONS, ANALYTICAL CHEMISTRY 66: 2318 (1994).
GANDHI, C.R., HEPATIC-EFFECTS OF ENDOTHELIN - RECEPTOR CHARACTERIZATION AND ENDOTHELIN-INDUCED SIGNAL TRANSDUCTION IN HEPATOCYTES, BIOCHEMICAL JOURNAL 287: 897 (1992).
GIAEVER, I, ANTIBODY-ANTIGEN REACTION - VISUAL OBSERVATION, JOURNAL OF IMMUNOLOGY 110: 1424 (1973).
GREANEY, M.D., IMPROVED METHOD FOR THE ROUTINE ANALYSIS OF ACETYLCHOLINE-RELEASE IN-VIVO - QUANTITATION IN THE PRESENCE AND ABSENCE OF ESTERASE INHIBITOR, JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS 622: 125 (1993).
Greene, L. A., Culturing Nerve Cells: 207 (1991).
GREENE, L.A., SYNTHESIS, STORAGE AND RELEASE OF ACETYLCHOLINE BY A NORADRENERGIC PHEOCHROMOCYTOMA CELL LINE, NATURE 268: 349 (1977).
JACKSON, T, STRUCTURE AND FUNCTION OF G-PROTEIN COUPLED RECEPTORS, PHARMACOLOGY & THERAPEUTICS 50: 425 (1991).
KIST, TBL, CAPILLARY ZONE ELECTROPHORESIS SEPARATION OF KININS USING A NOVEL LASER FLUORESCENCE DETECTOR, BRAZILIAN JOURNAL OF MEDICAL AND BIOLOGICAL RESEARCH 27: 11 (1994).
MONNIG, C.A., CAPILLARY ELECTROPHORESIS, ANALYTICAL CHEMISTRY 66: R280 (1994).
MORI, M, STRUCTURE OF CCF10, A PEPTIDE SEX-PHEROMONE WHICH INDUCES CONJUGATIVE TRANSFER OF THE STREPTOCOCCUS-FAECALIS TETRACYCLINE RESISTANCE PLASMID, PCF10, JOURNAL OF BIOLOGICAL CHEMISTRY 263: 14574 (1988).
NIE, S, unpublished data.
OATES, M.D., QUANTITATIVE AMINO-ACID-ANALYSIS OF INDIVIDUAL SNAIL NEURONS BY OPEN TUBULAR LIQUID-CHROMATOGRAPHY, ANALYTICAL CHEMISTRY 62: 1573 (1990).
OLEFIROWICZ, T.M., CAPILLARY ELECTROPHORESIS IN 2-MU-M AND 5-MU-M DIAMETER CAPILLARIES - APPLICATION TO CYTOPLASMIC ANALYSIS, ANALYTICAL CHEMISTRY 62: 1872 (1990).
PARCE, J.W., DETECTION OF CELL-AFFECTING AGENTS WITH A SILICON BIOSENSOR, SCIENCE 246: 243 (1989).
POZZAN, T, ACTIVATION OF MUSCARINIC RECEPTORS IN PC12 CELLS STIMULATION OF CA-2+ INFLUX AND REDISTRIBUTION, BIOCHEMICAL JOURNAL 234: 547 (1986).
ROGERS, K.R., ACETYLCHOLINE-RECEPTOR FIBER-OPTIC EVANESCENT FLUOROSENSOR, ANALYTICAL BIOCHEMISTRY 182: 353 (1989).
ROUNDS, M.A., ANION-EXCHANGE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH POSTCOLUMN DETECTION FOR THE ANALYSIS OF PHYTIC ACID AND OTHER INOSITOL PHOSPHATES, JOURNAL OF CHROMATOGRAPHY A 653: 148 (1993).
SANDS, S.B., CALCIUM PERMEABILITY OF NEURONAL NICOTINIC ACETYLCHOLINE-RECEPTOR CHANNELS IN PC12 CELLS, BRAIN RESEARCH 560: 38 (1991).
SMITH, R. D., ANALYTICAL CHEMISTRY 65: A574 (1993).
SUGAO, N, NA+/D-GLUCOSE COTRANSPORTER BASED BILAYER LIPID-MEMBRANE SENSOR FOR D-GLUCOSE, ANALYTICAL CHEMISTRY 65: 363 (1993).
SUZUKI, H, AN AMPEROMETRIC SENSOR FOR CARBON-DIOXIDE BASED ON IMMOBILIZED BACTERIA UTILIZING CARBON-DIOXIDE, ANALYTICA CHIMICA ACTA 199: 85 (1987).
Turner, A. P. F., Advances in Biosensors (1991).
UPDIKE, S.J., ENZYME ELECTRODE, NATURE 214: 986 (1967).
YAMAMOTO, K, J CHROMATOGR 588: 327 (1991).
