The tumor suppressor gene p53 is mutated in >50% of hepatocellular carcinomas (HCCs) in many geographic areas of the world, regardless of whether the HCC is associated with hepatitis B or C virus. It is not known whether the host produces antibodies to p53 in HCC; such antibodies have been detected in between 9% to 52% of patients with non-hepatic cancers, but in only 0.4% of healthy blood donors. These antibodies often appear prior to a diagnosis of cancer. Attempts were made to develop an assay to detect antibodies to p53 in HCC patients. An EIA was attempted by using p53 proteins produced by a recombinant vaccinia virus containing the gene for human p53. This was not successful because of a persistently weak signal in the assay. Subsequent approaches used the western blot method. p53 proteins were separated on SDS/PAGE and blotted onto a membrane, which was subsequently cut into narrow strips. Serum from each patient was diluted and incubated with each membrane strip and the reactive serum was identified by exposing the ECL-treated strip to X-ray film. Thirty-six serum samples from HCC patients and controls have been tested multiple times to develop reliable and reproducible conditions for the assay.

Agency
National Institute of Health (NIH)
Institute
Food and Drug Administration (FDA)
Type
Intramural Research (Z01)
Project #
1Z01BP004014-02
Application #
6161362
Study Section
Special Emphasis Panel (LOH)
Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Bureau of Health Planning and Resources Development
Department
Type
DUNS #
City
State
Country
United States
Zip Code