reactivation. immunoassay system), strips are impregnated with the dry conjugate , antibody, apo-enzyme, glucose and reagents for detecting hydrogen peroxide. The use of antiserum to glucose oxidase in the apoenzyme reactivation immunoassay system (ARIS) is described. Formation of an immune complex between. Apoenzyme reactivation immunoassay; Cofaetor-labeled; Inhibitor-labeled. Introduction. Homo~’:icous immunoassay is defined as an immunoassay system in.

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apoenzmye This application is also a continuation in part of, and claims the priority benefit of, copending U. Description of the Related Art There is a wide range of chemical entities test ligands, test analytes where rapid identification of the presence and relative levels of the entity are highly important.

The missing test ingredients—a regeneration buffer and a resuply of amplification substrate for the electrochemically active enzyme, will often be supplied by natural physiological process in the human or animals blood circulation or interstitial fluid. Test or analyte immunoassqy 19 acts to liberate prosthetic group 16 from a bound state to a more freely migrating state by the hydrolytic cleaving activity of test apoenzymme analyte enzyme Flavin adenine dinucleotide-labeled conjugates for use in specific binding assays.

Homogeneous apoenzyme reactivation immunoassay for thyroxin-binding globulin in serum.

The electrochemical apoenzyme reactivation technology of the present disclosure apoenztme be readily adapted to produce a prothrombin time test, other type of blood coagulation test, or other type of protease or hydrolytic enzyme cascade test.

Generally it will be advantageous to incorporate the hybrid antibody-apoenzyme genes into phages, and produce large number of variant versions of the hybrid antibody molecule, directed against different test antigens of interest, using phage display methods. By contrast, the work of Heller and others has shown that much higher efficiency can immunoasay obtained if the enzyme and electron transport mediator are both affixed to the electrode surface, and electrons can flow directly between the enzyme reaction center and the electrode by an electron transport mediator that is continuously attached to both the enzyme and the electrode surface.


In the example where a prothrombin time coagulation test is desired, a coagulation immuoassay, such as a thromboplastin-calcium solution, is made up and a small approximately ul drop of this solution is applied to a plastic cover.

Other immunochemical applications for this technology include tests for sepsis, angiogenesis, pregnancy and ovulation, cardiovascular status, infectious disease, drugs of abuse, therapeutic drugs, kidney disease, ischemia, and cancer diagnostics. Apoenzyme 24 is reconstituted to an active enzyme 24 configuration, and the active site 25 of enzyme 24 changes from an inactive configuration to an active configuration. As a result, addition of excess FAD groups from the test analyte detection moieties will have no effect on enzyme activity.

FAD can be coupled to peptides and proteins by the methods of Schroeder et. The thromboplastin activates immunoasday VII, which in turn activates factor X, which in turn activates thrombin, which in turn converts fibrinogen to fibrin, forming a clot.

This combination creates a novel combination test technology capable of detecting a wide range of different analytes, reactivatiin operating in a wide variety of wet or dry, in vivo or in vitro environments.

Pregnancy and ovulation markers include early pregnancy factor, human chorionic gonadotropin and luteinising hormone LH. This method also relies on directly detecting the concentration of the analyte e.

Meaning of “apoenzyme” in the English dictionary

This distinction is critical. One of the more modern, and particularly favored, methods is according to the methods of Heiss et. The present application is focused on this latter type of rapid chemical test methods. Creatinine biosensor based on ammonium ion selective electrode apoemzyme its application in flow-injection analysis. If use with fresh whole blood is desired, no other components need be present. In some cases, it may be desirable to incorporate the device into the flow cell devices previously described in Ser.

This reaction liberates electrons 52which can flow, by way of electron transport mediator 53 to the electron transporting zones 54 of porous electrode Reactivaton upon the specifics of the experiment, various deposition patterns can be used. The rfactivation device additionally contains thromboplastin 9which is used to initiate the extrinsic coagulation pathway that leads to blood coagulation. This is shown in FIG. The blocker part of the analyte detection moiety is an entity that, in the absence of interactions between the moiety’s detector region and the test ligand, acts to prevent the enzyme activation factor from binding to the inactivated enzyme apoenzyme on the sensing electrode.


Meaning of “apoenzyme” in the English dictionary. This is because in many cases, simple and practical ways to transduce the chemical signal produced by the test reagent-test analyte reaction over to an electrochemical signal capable of being reactivatoon at a test reagent readtivation has not been identified.

Homogeneous apoenzyme reactivation immunoassay for thyroxin-binding globulin in serum.

The test strip is attached to the electrochemistry measuring apparatus. Note that because the hybrid antibody-apoenzyme 1221 is connected to the support 313by a flexible tether 414the test device may be washed or flushed with fresh reaction buffer normally containing salts, a pH control buffer, and sufficient quantities of the amplification substrate for the electrically active enzyme to provide an adequate electrochemical signal for the next assay.

Prussian blue modified amperometric FIA biosensor: This is because such analyte enzymes usually do not create apoenzyme prosthetic groups as a reaction product, which is required by A’s teaching.

SUMMARY OF THE INVENTION The invention discloses methods in which dry reagent electrochemical technology, which is in a relatively mature form due to the extensive amount of development pioneered by the blood glucose monitoring industry, may be simply adapted to perform important hydrolase enzymatic activity tests such as blood coagulation and other biological tests of interest.

In use, a patient test sample 10 such as blood or plasma is applied to the test device.