Receptor-based histamine assay
AssigneeMinnesota Mining and Manufacturing Company
InventorsThomas K. Rice, Therese A. Senta
Life of this patent
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Abstract
A receptor-based clinical or research competitive inhibition assay method for determining histamine in body or laboratory fluids is disclosed. A kit for carrying out the method is also described comprising an amount of histamine receptors sufficient to provide an excess of receptors for binding with the amount of histamine in the sample to be assayed, and an amount of a histamine-indicator conjugate sufficient to react with the resulting unbound histamine receptors present after reaction of the histamine sample with the histamine receptors.
Claims16
A method for determining the amount of histamine in a liquid test sample using histamine receptors and a histamine-indicator conjugate comprising the steps of:
- a. contacting a measured amount of said test sample with an excess of substantially Purified histamine receptors which are free in solution or bound to a continuous solid phase support,
- b. incubating and reacting said test sample with said histamine receptors, which reaction leaves unbound histamine receptors available for further bonding,
- c. contacting an amount of histamine-indicator conjugate with the bound and unbound histamine receptors sufficient to bind said conjugate to all unbound histamine receptors,
- d. incubating and reacting said histamine-indicator conjugate with said unbound histamine receptors,
- e. removing unbound histamine-indicator conjugate if necessary to measure the amount of histamine-indicator conjugate bound to said histamine receptors,
- f. measuring the amount of histamine-indicator conjugate bound to the histamine receptors, and
- g. determining the amount of histamine in said test sample by comparison with standard curves,
- with the proviso that
- said histamine-indicator conjugate maintains its activity or undergoes stimulation of activity when bound to said histamine receptors, and
- said method being capable of measuring at least 0.05 ng of histamine in a test sample.
A method for determining the amount of histamine in a liquid test sample using histamine receptors and histamine-alkaline phosphates as a histamine-indicator conjugate comprising the steps of:
- a. contacting a measured amount of said test sample with an excess of substantially purified histamine receptors which are free in solution or bound to a continuous solid phase support,
- b. incubating and reacting said test sample with said histamine receptors, which reaction leaves unbound histamine receptors available for further bonding,
- c. contacting an amount of histamine-alkaline phosphatase with the bound and unbound histamine receptors sufficient to bind said conjugate to all unbound histamine receptors,
- d. incubating and reacting said histamine-alkaline phosphatase with said unbound histamine receptors,
- e. removing unbound histamine-alkaline phosphatase if necessary to measure the amount of histamine-alkaline phosphatase bound to said histamine receptors,
- f. measuring the amount of histamine-alkaline phosphatase bound to the histamine receptors, and
- g. determining the amount of histamine in said test sample by comparison with standard curves,
- with the proviso that
- said histamine-alkaline phosphatase undergoes stimulation of activity when bound to said histamine receptors, and
- said method being capable of measuring at least 0.05 ng of histamine in a test sample.
Dependencies are inferred from the claim text.