Buffered wash composition, insolubilizing composition, test kits and method of use
AssigneeEastman Kodak Company
InventorsGregory J. McClune, Karen L. Findling
Life of this patent
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Abstract
A buffered aqueous composition is useful simultaneously as a wash solution and a dye-providing composition in specific binding assays involving enzyme-labeled specific binding reagents. The wash composition includes a dye-providing composition, a buffer and an organic solvent having a certain molecular weight and water-solubility. Another useful composition includes a particulate substrate having avidin attached thereto, and a peroxidase reducing agent. Either composition can be provided in a diagnostic test kit, and can be used to detect a specific binding ligand in assays.
Claims10
A method for the determination of a specific binding ligand comprising the steps of:
- A. contacting a specimen suspected of containing said specific binding ligand with an enzyme-labeled receptor for said specific binding ligand to form a detectable complex of ligand and receptor,
- B. separating uncomplexed receptor from said detectable complex by washing said detectable complex with a buffered aqueous wash composition comprising:
- a. a composition which provides a dye in response to said enzyme,
- b. a buffer, and
- c. a water-soluble organic solvent which has a molecular weight between about 40 and about 100 and is present in an amount of from about 2.5 to about 25 volume %, and
- C. detecting the resulting dye as an indication of the presence of said specific binding ligand in said specimen.
A method for the detection of human chorionic gonadotropin comprising the steps of:
- A. contacting a specimen suspected of containing human chorionic gonadotropin with a first antibody directed to hCG to form an immunological complex,
- B. simultaneously or subsequently to step A, contacting said specimen with a second antibody to hCG to form a sandwich complex, said first and second antibodies specifically binding to different epitopes, and at least one of said antibodies being labeled with peroxidase, and the other antibody either insolubilized or capable of becoming so,
- C. separating uncomplexed materials from said sandwich complex through a filtration membrane by washing said sandwich complex with a buffered wash composition while in the presence of hydrogen peroxide, said wash composition comprising:
- a. a composition which provides a dye in response to peroxidase,
- b. a buffer, and
- c. a water-soluble organic solvent which has a molecular weight between about 40 and about 100 and is present in an amount of from about 2.5 to about 25 volume %, and
- D. detecting the resulting dye on said membrane as an indication of the presence of hCG in said specimen.
Dependencies are inferred from the claim text.