Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
1999-02-14
2001-06-26
Huff, Sheela (Department: 1642)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S004000, C435S007920, C435S007940, C530S387100
Reexamination Certificate
active
06251613
ABSTRACT:
TECHNICAL FIELD
The present invention relates to a method of detecting an anti-GADII antibody, and a method of diagnosing cancer using this detecting method. In particular, it relates to a method of diagnosing cancer by detecting an anti-GADII antibody existing in serum.
BACKGROUND ART
Cancer is known to occur because of some abnormality in a gene. A change or abnormality in the gene at its transcription level, in particular, is regarded as a major cause of cancer (“Science” Vol. 222, 1983, pp. 765-771). To elucidate the mechanism of onset of cancer, the acquisition of a protein variedly expressed during carcinogenesis and a gene encoding the protein, or a protein different in the state of expression among tissues and a gene encoding the protein has been performed eagerly since about 1980. For example, &agr;-fetoprotein and CEA (carcinoembryonic antigen) are known as cancer-specific proteins which were acquired by biochemically analyzing a carcinomatous tissue, and searching for its difference from a normal tissue.
Nevertheless, only a part of carcinogenesis mechanism has been clarified so far, thus leaving a demand for further elucidation of carcinogenesis-related genes and proteins. Also demanded is a novel method of diagnosing cancer using these carcinogenesis-related genes and proteins.
DISCLOSURE OF THE INVENTION
In order to isolate and elucidate a novel protein whose expression increases during the process of carcinogenesis and a novel gene encoding this protein, the inventors extracted genes specifically expressed in rat hepatic cancer by using subtraction method, and isolated the genes whose expression increased in hepatic cancer according to analysis using dot-screening method. The full length cDNA's of the genes were obtained from a hepatic cancer cDNA library, and the base sequences of the genes were determined. Further, amino acid sequences encoded by the genes were determined.
Moreover, Northern blot hybridization confirmed the cDNA's to be hepatic cancer specific genes. Thus, inventors isolated rat and human GADII genes which were novel genes specific to hepatic cancer. Also, the inventors produced rat GADII protein in
E. coli,
inoculated this protein into a mammal other than a species from which the protein was derived excluding human, to prepare antibodies against the protein and confirm the antigenicity of the protein.
The present invention provides a method of detecting an antibody against the above-mentioned cancer-specific protein, and a method of diagnosing cancer using this detecting method.
The inventors have found that the antibodies against GADII protein which was a cancer-specific protein already invented by the inventors are remarkably detected in a living organism upon carcinogenesis, thereby accomplishing the present invention.
The present invention provides a method of easily screening the onset of cancer by using GADII, which is a protein remarkably expressed in cancer, such as hepatic cancer in particular, so as to detect an antibody against this protein.
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Baekkeskov, et al., “Identificiation of the 64K autoantigen in insulin-dependent diabetes as the GABA-synthesizing enzyme glutamic acid decarboxylase” Nature, vol. 347, Sep. 13, 1990, pp. 151-156.
Michelsen, et al., “Cloning, characterization, and autoimmune recognition of rat islet glutamic acid decarboxylase in insulin-dependent diabetes mellitus” Proc. Natl. Acad. Sci. USA, vol. 88, Oct. 1991, pp 8754-8758.
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Kishimoto Toshihiko
Makino Yasutaka
Tamura Taka-aki
Helms Larry R.
Huff Sheela
Pillsbury & Winthrop LLP
Sumitomo Electric Industries Ltd.
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