| 摘要 |
A toxicoproteomic method for developing immunoassays to determine if a toxicant or a disease causes damage to a specific organ is disclosed. The discovery phase of the method involves the administration of a toxicant to an animal which will cause damage to a specific organ of the animal, which damage results in the production of an oxidatively modified, organ specific protein(s). Blood serum, which includes the modified protein(s) released from the organ following damage, is collected from the animal. The modified protein(s) is isolated from serum by immunoaffinity purification using antibodies directed against any number of damage-related, amino acid modifications, in this instance, nitrotyrosine, chlorotyrosine and methionine sulfoxide. Its function as a biomarker is validated by determining if the modified protein(s) is specific to the tissue or organ of interest. If the modified protein is organ specific, an antibody is raised against it, and this antibody is utilized in immunoassays to (1) measure the concentration of the protein in serum and, (2) together with antibodies against the modified amino acids found on its surface used to determine the type and extent of modification found on the protein. The assays of the present invention have utility for the toxicological screening of newly developed drugs, evaluating patients diagnostically or for organ involvement in disease, the study of cellular responses to various toxicants, and the determination of peripheral redox status.
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