Single cells as biosensors for chemical separations.
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.
Shear
J B
JB
Department of Chemistry, Stanford University, CA 94305.
Fishman
H A
HA
Allbritton
N L
NL
Garigan
D
D
Zare
R N
RN
Scheller
R H
RH
D000109:Acetylcholine / Q000032:analysis / Q000302:isolation & purification; D000255:Adenosine Triphosphate / Q000032:analysis / Q000302:isolation & purification; D000818:Animals; D015374:Biosensing Techniques; D001920:Bradykinin / Q000032:analysis / Q000302:isolation & purification; D002118:Calcium / Q000032:analysis; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D004586:Electrophoresis; D008024:Ligands; D008856:Microscopy, Fluorescence; D009865:Oocytes; D016716:PC12 Cells; D018408:Patch-Clamp Techniques; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000032:analysis / Q000302:isolation & purification; D015398:Signal Transduction; D014982:Xenopus laevis
D008024:Ligands; D012701:Serotonin; D000255:Adenosine Triphosphate; D000109:Acetylcholine; D001920:Bradykinin; D002118:Calcium