Detection of sub-nanogram quantities of Mojave toxin via enzyme immunoassay with light addressable potentiometric detector.
Sensitive detection of Mojave toxin (MoTX), a potent neurotoxin isolated from the venom of Crotalus scutulatus scutulatus, was developed using an enzyme immunoassay (EIA) and light addressable potentiometric detection. This EIA utilizes both biotin- and fluorescein-labeled anti-MoTX antibodies to immobilize and detect sub-nanogram to nanogram quantities of toxin. Labeled mono- and polyclonal anti-MoTX antibodies were used alone and/or in combination to determine maximum assay sensitivity. Assays performed using a combination of biotinylated poly- and fluoresceinated monoclonal antibodies produced an assay with a lower detection limit near 2.5 ng. Assays performed using labeled polyclonals alone, or in combination with biotinylated mono- and fluoresceinated polyclonal antibodies, indicated increased sensitivity with a detection limit near 300 pg. In conclusion, we describe an enzyme immunoassay using different labeled antibody schemes which detects sub-nanogram quantities of MoTX via light addressable potentiometric detection.
Colston
J T
JT
Brain Research Laboratory of Biochemistry, University of Texas, San Antonio 78249.
Kumar
P
P
Rael
E D
ED
Tsin
A T
AT
Valdes
J J
JJ
Chambers
J P
JP
D000906:Antibodies; D000911:Antibodies, Monoclonal; D015374:Biosensing Techniques; D003435:Crotalid Venoms / Q000032:analysis*; D007124:Immunoenzyme Techniques; D007202:Indicators and Reagents; D008832:Microchemistry; D009498:Neurotoxins / Q000032:analysis*; D011199:Potentiometry / Q000379:methods
D000906:Antibodies; D000911:Antibodies, Monoclonal; D003435:Crotalid Venoms; D007202:Indicators and Reagents; C018107:Mojave toxin; D009498:Neurotoxins