An immunoassay for terbutryn using direct hapten linkage to a glutaraldehyde network on the polystyrene surface of standard microtiter plates.
2-Aminobutylamino-4-ethylamino-6-isopropylamino-1,3,5-triazine (ABA-atrazine) has been synthesized and used as a coating hapten in an immunoassay with a monoclonal antibody against terbutryn. Coating was achieved by covalently linking ABA-atrazine to a glutaraldehyde polymer network directly bound to the polystyrene surface of a standard 96-well microtiter plate. The assay was carefully optimized. In particular, the coating hapten concentration had a strong effect on the ELISA sensitivity. By including a pre-incubation step a low test midpoint (IC50-value) of 0.130 microg L(-1) was achieved. As far as we are aware this is the most sensitive ELISA for terbutryn yet reported. The coating-hapten-format presented is proposed as generally applicable, because the glutaraldehyde-modified microtiter plate surface enables stable immobilization of a broad variety of coating haptens.
Holthues
H
H
Institute of Physical Chemistry, University of Mainz, Germany.
Pfeifer-Fukumura
U
U
Hartmann
I
I
Baumann
W
W
D000911:Antibodies, Monoclonal / Q000276:immunology; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D005976:Glutaral / Q000737:chemistry; D006241:Haptens / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D006540:Herbicides / Q000032:analysis* / Q000276:immunology; D011108:Polymers / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry; D012680:Sensitivity and Specificity; D013499:Surface Properties; D014227:Triazines / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D006241:Haptens; D006540:Herbicides; D011108:Polymers; D011137:Polystyrenes; D014227:Triazines; D005976:Glutaral; C010347:terbutryne