Detection of digitalis compounds using a surface plasmon resonance-based biosensor.
An automated surface plasmon resonance-based biosensor system has been used to detect endogenous and exogenous digitalis-like factors (EDLF) in the pmolar range in real time. EDLF was purified from umbilical cord blood. EDLF has been suggested to play a role in hypertension and in perinatal adaptation. Highly specific polyclonal anti-ouabain antibodies showed a high affinity binding capacity for ouabain, ouabagenin and strophantidin with an IC50 value of 5 x 10(-10) M, 7.0 x 10(-10) M and 2 x 10(-8) M, respectively. EDLF cross-reacted with antibodies and its concentration in plasma at IC50 was around 50 pmol ouabain equivalent. This study shows the potential usefulness of the biosensor technology for biomolecular interaction analysis. The features of this technology (fully automated, measured in real time, sharpened response) offer several advantages compared with a traditional immunoassay like radioimmunoassay (RIA) in the detection of digitalis compounds in human fluids.
Laricchia-Robbio
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Institute of Mutagenesis and Differentiation, CNR, Pisa, Italy.
Balzan
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Ghione
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Montali
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Revoltella
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RP
D000906:Antibodies / Q000097:blood; D002298:Cardenolides; D004077:Digoxin; D004791:Enzyme Inhibitors / Q000032:analysis*; D006801:Humans; D007231:Infant, Newborn; D010042:Ouabain / Q000276:immunology; D011863:Radioimmunoassay; D012503:Saponins / Q000032:analysis*; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors*; D020349:Surface Plasmon Resonance
D000906:Antibodies; D002298:Cardenolides; D004791:Enzyme Inhibitors; D012503:Saponins; C034504:digoxin-like factors; D010042:Ouabain; D004077:Digoxin; D000254:Sodium-Potassium-Exchanging ATPase