Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1991-09-16
1994-03-29
Hill, Jr., Robert J.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 74, 435 792, 435 794, 5303871, 436548, G01N 3353
Patent
active
052983935
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a monoclonal antibody which recognizes glutathione S-transferase (hereinafter abbreviated as GST) that can be used as the diagnostic reagent for carcinomas such as cancer of the liver, stomach cancer, etc. and a process for preparing it.
BACKGROUND OF THE ART
GST is known as a detoxicating enzyme which catalyzes the conjugation (reaction) of various substances, which enter into an organism, or analogous substances (hereinafter referred to as substrates) which are produced in vivo, with glutathione of reduced type. Many of the substrates react with the in vivo protein or nucleic acid to cause a change to a morbid state (for instance, carcinogenesis) but the reactivity of these substrates are neutralized by said conjugation to be transformed into more water-soluble products and metabolized in the liver, etc. and finally excreted out of the body.
GST is found existing in various kinds of living species inclusive of mammals and is especially contained in considerable quantities in the liver, spleen, kidney, lung, brain, skeletal muscles, placenta, and small intestine, and also found in the skin, red blood cell, and white blood corpuscule though very small in amount. GST generally comprises various types of enzymes and is not only specific to species but also specific to the internal organs and tissues.
In the case of a human being, basic GST and acid GST are known (see The Journal of Biological Chemistry, vol. 259, No. 20, pp. 12,444-12,448 and pp. 12,449-12,455 (1984)). Basic GST has the isoelectric point (pI) of 7-9 and consists of two subunits, each subunit having a molecular weight of about 23,000. Acid GST has the isoelectric point (pI) of 4-5 and consists of two subunits, each subunit having a molecular weight of about 22,000. Basic GST exists mainly in the liver of a healthy normal adult and also in other places such as kidney, testicle, small intestine, brain, and lung though less in amount. While acid GST scarcely exists in the liver of a healthy normal adult but found existing in the liver of a newborn baby, placenta, such proliferous cells as liver cancer cells, stomach cancer cells, etc., red blood cell, and white blood corpuscule. Since acid GST produced by the proliferous cells is extricated out of the site of its production into the blood, it has possibilities of being utilized as a tumor marker highly specific to such digestive system cancers as liver cancer, stomach cancer, colon cancer, esophagus cancer, and cancer of the pancreas, and such blood tumors as cancer of the blood and lymphocytoma.
DISCLOSURE OF THE INVENTION
The present inventors have conducted an intensive research to obtain a monoclonal antibody which can detect the aforementioned tumor marker with high accuracy and finally prefected the present invention.
This invention is concerned with monoclonal antibody which recognizes human acid GST. Though it is naturally presumed that human acid GST somewhat differs from each other in structure depending upon the internal organ or tissue where it is produced, it is acid GST produced from the liver of a newborn baby, placenta, red blood cell, and proliferous cells such as liver cancer cell and stomach cancer cell that are useful as the marker to be used for the diagnosis of cancer. Therefore, a monoclonal antibody which recognizes such acid GST in common (for instance, which recognizes common subunits) should desirably be used in the present invention, and more particularly a mouse antibody of IgG type is desirable. Also, the monoclonal antibody of the present invention has a characteristic property that it does not exhibit a cross reaction with human basic GST.
The present invention also includes a method for determing acid GST with the use of the aforementioned monoclonal antibody and a kit of reagents to be used such determination.
BRIEF DESCRIPTION OF DRAWING
FIG. 1 is a drawing to show the relation between the concentration of acid GST and the optical density (420 nm).
BEST MODE OF CARRYING OUT THE INVENTION
The monoclon
REFERENCES:
patent: 4486530 (1984-12-01), David et al.
Kohler, et al., Nature vol. 256, "Continuous Cultures of Fused Cells Secreting Antibody of Predefined Specificity", pp. 495-497, Aug. 7, 1975.
Maruyama et al., The Journal of Biological Chemistry, vol. 259, No. 20, Oct. 25, 1984, 12444-12448, "Distinctions between the Multiple Cationic Forms of Rat Liver Glutathione S-transferase".
Wang et al., Different Forms of Ya Subunit of Ligandin and Glutathione S-Transferase B Detected by Monoclonal Antibodies, Mar. 1, 1985, Federation Proceedings, vol. 44, No. 3, p. 861.
Chemical Abstracts, vol. 100, No. 11, Mar. 12, 1984 p. 406, Abstract No. 83937t.
Honda Hitomi
Hosoda Kenji
Maruyama Hiroshi
Masuho Yasuhiko
Niitsu Yoshiro
Hill Jr. Robert J.
Scheiner Laurie
Teijin Limited
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