Facets and formulations of enzymes, immunoglobulins, and receptors for assay and sensor applications

A study on the application of enzymes, immunoglobulins and receptors for assays and sensors. Enzymes were exploited in the detection of an inhibitor and for the simultaneous determination of multiple substrates. Immunosensors with potentiometric and amperometric detection were explored. The amperometric detection of tricyclic antidepressants and a microtiter plate based assay using the acetylcholine receptor are also described.;Detection of an enzyme inhibitor by an enzyme sensor was demonstrated by detecting allopurinol with a carbon paste electrode coated with xanthine oxidase. This project provided insight into some of the considerations involved with this type of analysis such as enzyme loading, reversibility and the mechanism of interaction between the enzyme and the inhibitor.;Multicomponent analysis was achieved by utilizing an array of enzymes for the detection of their respective substrates. The enzyme array consisted of glucose oxidase and glucose-6-phosphate dehydrogenase immobilized onto polydextran beads derivatized with polyornithine. A charge coupled device (CCD) was employed to record the response of the array and image analysis allowed responses of the individual substrates to be spatially resolved.;Potentiometric ionophore mediated immunoassays (PIMIA) using aminated hydrophobic compounds as ionophoric haptens were investigated. The potential produced relied upon the aminated compounds acting as ionophores for protons.;Immunoassays based on the use of enzyme modulator(coformycin)-hapten conjugates and enzyme(glucose oxidase)-hapten conjugates were investigation. Coformycin conjugates to digoxin and a benzodiazepine served as model systems for assays based on enzyme modulators. Preliminary studies did show that this inhibitor can be conjugated and retain biological function. Studies of an immunosensor model using three enzymes, glucose oxidase, peroxidase and catalase produced an indication that an amperometric immunosensor based on the spatial discrimination of hydrogen peroxide generated from glucose and oxygen could be possible.