Detection and quantitation of 8-OH-Adenine using monoclonal antibodies
AssigneeCytoChem, Inc.
InventorsEric H. Holmes, Thomas G. Greene
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Abstract
The present invention relates generally to assays for 8-hydroxyadenine (8-OH-Ade) and, more particularly, to improved immunoassays for the detection and quantitation of 8-OH-Ade in biological specimens. The present invention further relates to new monoclonal antibodies directed against 8-OH-Ade for use in the immunoassays.
Claims36
A monoclonal antibody obtained from a hybridoma selected from the group consisting of hybridomas ATCC HB12188 and ATCC HB12189.
A hybridoma selected from the group consisting of hybridomas ATCC HB12188 and ATCC HB12189.
A method for determining the presence or amount of 8-OH-adenine in a biological specimen comprising the steps of contacting the specimen with a monoclonal antibody obtained from a hybridoma selected from the group consisting of hyridomas ATCC HB12188 and ATCC HB12189 to form a complex of monoclonal antibody and 8-OH-adenine, and determining the presence or amount of the complex formed as an indication of the presence or amount of 8-OH-adenine in the specimen.
A method for diagnosing or monitoring the status of a disease associated with elevated levels of 8-OH-adenine in a human or animal comprising the steps of:
- a. obtaining a biological specimen of nucleic acids from the human or animal;
- b. contacting the specimen with a monoclonal antibody obtained from a hybridoma selected from the group consisting of hyridomas ATCC HB12188 and ATCC HB12189 to form a complex of monoclonal antibody and 8-OH-adenine;
- c. determining the amount of complex formed as a measure of the presence or amount of 8-OH-adenine in the specimen wherein the amount of complex determined is indicative of a disease state associated with elevated levels of 8-OH-adenine or is correlated with the status of a disease associated with elevated levels of 8-OH-adenine.
A method for determining exposure of a human or animal to a toxicant associated with elevated levels of 8-OH-adenine in a biological specimen obtained from the human or animal comprising the steps of:
- a. obtaining the biological specimen from the human or animal;
- b. contacting the specimen with a first amount of monoclonal antibody obtained from a hybridoma selected from the group consisting of hyridomas ATCC HB12188 and ATCC HB12189 to form a complex of monoclonal antibody and 8-OH-adenine;
- c. determining the amount of complex formed in step b by using an 8-OH-adenine standard curve, the standard curve obtained by the method of contacting a second amount of monoclonal antibody to a known amount of 8-OH-adenine;
- and
- d. comparing the amount of complex determined in step c with a range of 8-OH-adenine or 8-OH-deoxyadenine concentrations found in human or animal specimens that have not been exposed to a toxicant associated with elevated levels of 8-OH-adenine.
A method for determining the amount of 8-OH-adenine or 8-OH-deoxyadenine in a biological specimen comprising:
- a. contacting a monoclonal antibody obtained from a hybridoma selected from the group consisting of hybridomas ATCC HB12188 and ATCC HB12189 with the specimen to form a complex between the antibody and 8-OH-adenine in the specimen;
- b. quantitating the amount of complex formed;
- and
- c. comparing the amount of complex formed to the amount of the monoclonal antibody that complexes with a known amount of 8-OH-adenine or 8-OH-deoxyadenosine.
A method for determining the presence or amount of 8-OH-adenine in a biological specimen comprising the steps of:
- a. contacting the specimen with a monoclonal antibody obtained from a hybridoma selected from the group consisting of hybridomas ATCC HB12188 and ATCC HB12189 to form a first complex comprising the monoclonal antibody and 8-OH-adenine;
- b. contacting the specimen containing the first complex with a second antibody capable of binding to the first complex;
- c. determining the amount of second complex formed as a measure of the presence or amount of 8-OH-adenine in the specimen.
A method for determining the presence or amount of an epitope which comprises 8-OH-adenine in a biological specimen comprising the steps of:
- a. contacting the specimen with a monoclonal antibody obtained from a hybridoma selected from the group consisting of hybridomas ATCC HB12188 or ATCC HB12189 to form a complex of monoclonal antibody and epitope, and
- b. determining the presence or amount of complex formed as a measure of the presence on amount of epitope in the specimen.
Dependencies are inferred from the claim text.