Effects of receptor concentration, media pH and storage on nicotinic receptor-transmitted signal in a fiber-optic biosensor.
The optical signal generated by a nicotinic acetylcholine receptor (nAChR)-based optical biosensor was dependent on the density of toxin binding sites (i.e. receptors) immobilized on the surface of the fiber. The maximum density of nAChR receptors absorbed on the optic fibers, measured using [125I]-alpha-bungarotoxin (the alpha-neurotoxin of the Bungarus snake venom) binding, was 6.2 pmol of receptor sites per fiber. Incubation time, that was required for maximal noncovalent immobilization of the receptor protein on the fiber, was less than 10 min. Immobilization of the nAChR protein on the quartz fiber was affected by pH of the medium, with pH 3.5 as the optimal. Stored in phosphate buffered saline (PBS) or in a dry nitrogen atmosphere, the nAChR optical sensor showed no loss of activity over the first 3 days, then showed a slow but gradual loss in activity (45% over the next 30 days).
Rogers
K R
KR
Department of Pharmacology and Experimental Therapeutics, University of Maryland School of Medicine, Baltimore 21201.
Valdes
J J
JJ
Eldefrawi
M E
ME
D000818:Animals; D015374:Biosensing Techniques; D002038:Bungarotoxins / Q000378:metabolism; D005336:Fiber Optic Technology; D019793:Fluorescein; D005452:Fluoresceins; D005470:Fluorometry; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D055100:Optical Fibers; D011978:Receptors, Nicotinic / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*
D002038:Bungarotoxins; D005452:Fluoresceins; D007457:Iodine Radioisotopes; D011978:Receptors, Nicotinic; D019793:Fluorescein