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Method for simultaneous determination of enzymatic activities of enzymes

  • US 3,925,162 A
  • Filed: 08/06/1973
  • Issued: 12/09/1975
  • Est. Priority Date: 08/17/1972
  • Status: Expired due to Term
First Claim
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1. A METHOD FOR THE SIMULTANEOUS DETERMINATION OF ENZYMATIC ACTIVITIES OF A PLURALITY OF ENZYMES INA SINGLE REACTION MEDIUM, COMPRISING:

  • ADDING A SUBSTRATE FOR EACH OF THE ENZYMES TO BE DETERMINED, EACH SUBSTRATE BEING SELECTED SUCH THAT IT IS CAPABLE OF GIVING AN ENZYMATIC REACTION WITH ITS RESPECTIVE ENZYME, IT DOES NOT INTERACT WITH THE OTHER SUBSTRATES REACTION PRODUCTS OR REAGENTS USED AND IT DOES NOT ADVERSELY AFFECT THE MEASUREMENT OF THE ENZYMATIC ACTIVITY, TO AN AQUEOUS SOLUTION CONTAINING A PLURAITY OF ENZYMES TO BE DETERMINED, THE ENZYMES BEING SELECTED SUCH THAT THE ENYZMATIC CONDITIONS OF EACH ARE SIMILAR TO EACH OTHER AND THEY CAN BE REACTED WITH THEIR RESPECTIVE SUBSTRATES UNDER THE SAME REACTION CONDITIONS;

    SIMULTANEOUSLY REACTING EACH OF THE ENZYMES WITH THEIR RESPECTIVE SUBSTRATES UNDER THE OPTIMUM REACTION CONDITIONS WITH RESPECT TO EACH OF THE ENZYMES UNDER WHICH THE ENZYMATIC REACTIONS OF THE PLURALITY OF ENZYMES TO BE DETERMINED TAKE PLACE SIMULTANEOUSLY;

    MEASURING SIMULTANEOUS CHANGES IN THE ABSORBANCE OR FLUORESCENCE OF THE RESULTING REACTION SYSTEM WITH THE LAPSE OF TIME AT A PLUTALITY OF OPTIONAL WAVELENGTHS WHICH ARE DIFFERENT FROM EACH OTHER AND THE NUMBER OF WHICH IS EQUAL TO THE NUMBER OF ENZYMES TO BE DETERMINED;

    FORMULATING SIMULTANEOUS EQUATIONS OF THE FIRST DEGREE BY TAKING ADVANCING OF THE PROPOTIONAL RELATIONSHIP, WHICH IS INDEPENDENT WITH RESPECT TO EACH OT THE ENZYMES, BETWEEN THE MEASUREMENT VALUES AND THE ENZYMATIC ACTIVITIES;

    AND DETERMINING THE ENZYMATIC ACTIVITY OF EACH OF THE ENZYMES BY THE THUS FORMULATED EQUATIONS.

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