Claims for Patent: 5,175,084

Summary for Patent: 5,175,084

Title:	Method for the diagnosis of hepatic carcinoma
Abstract:	A method for diagnosing hepatic carcinoma, which is characterized by enzyme-immunologically measuring the amount of collagenase inhibitor present in sera, plasmas of synovial fluids by way of a sandwich assay wherein two different monoclonal antibodies which specifically bind to different antigenic determinants of the collagenase inhibitor are used, and comparing the measured amount with that for normal subjects.
Inventor(s):	Inoue; Kyoichi (Toyama, JP), Ichida; Takafumi (Toyama, JP), Miyagiwa; Miki (Toyama, JP), Hayakawa; Taro (Nagoya, JP), Kodama; Shuji (Takaoka, JP), Iwata; Kazushi (Takaoka, JP)
Assignee:	Fuji Yakuhin Kogyo Kabushiki Kaisha (Toyama, JP)
Application Number:	07/382,789
Patent Claims:	1. A screening method to aid in the diagnosis of hepatic carcinoma, comprising: (a) contacting a sample of serum or plasma suspected of containing collagenase inhibitor with a first insoluble complex composed of a solid substrate having bound thereto a first monoclonal antibody specific for collagenase inhibitor in order to bind said collagenase inhibitor to said insoluble complex; (b) contacting said insoluble complex with a second monoclonal antibody specific for collagenase inhibitor conjugated to an enzyme so as to form an insoluble ternary complex of said enzyme-conjugated second monoclonal antibody, said collagenase inhibitor, and said solid substrate; (c) determining the amount of said collagenase inhibitor by incubating said ternary complex with a substrate of said enzyme and determining the amount of said enzyme reaction product formed from a standard curve to provide a measure of the amount of collagenase inhibitor; and (d) comparing the amount of said collagenase inhibitor to that in serum or plasma from normal subjects, wherein an elevation of said amount of collagenase inhibitor is indicative of the presence of hepatic carcinoma. 2. The method of claim 1, wherein said solid substrate is a material selected from the group consisting of polystyrene, polycarbonate, polypropylene, and polyvinyl. 3. The method of claim 1, wherein said solid substrate is in a form selected from the group consisting of a ball, a microplate, a stick, and a test tube. 4. The method of claim 1, wherein said solid substrate is a polystyrene ball. 5. The method of claim 1, wherein said enzyme is horseradish peroxidase. 6. The method of claim 5, wherein said substrate is a substrate for horseradish peroxidase. 7. The method of claim 5, wherein said amount of said collagenase inhibitor is determined by measuring the optical density of the reaction mixture containing said enzyme reaction product formed at 450 nm. 8. The method of claim 1, wherein said standard curve is prepared by measuring the amount of enzyme reaction product formed in reaction mixtures containing varying amounts of human serum collagenase inhibitor. 9. The method of claim 1, wherein said screening further comprises performing liver function tests. 10. The method of claim 11, wherein said liver function tests are selected from the group consisting of the zinc sulfate turbidity test, measurement of glutamic oxaloacetic transaminase, measurement of glutamic-pyruvic transaminase, measurement of alkaline phosphatase, measurement of lactate dehydrogenase, measurement of .gamma.-glutamyl transpeptidase, measurement of prolyl hydroxylase, and measurement of .alpha.-fetoprotein. 11. A screening method to aid in the diagnosis of hepatic carcinoma, comprising: (a) contacting a sample of serum or plasma suspected of containing collagenase inhibitor with a first insoluble complex comprised of a solid substrate having bound thereto an antibody produced by hybridoma clone FERM BP-3469 in order to bind said collagenase inhibitor to said insoluble complex; (b) contacting said insoluble complex with a second monoclonal antibody produced by hybridoma clone FERM BP-3468 conjugated to an enzyme so as to form an insoluble ternary complex of said enzyme-conjugated second monoclonal antibody, said collagenase inhibitor, and said solid substrate; (c) determining the amount of said collagenase inhibitor by incubating said ternary complex with a substrate of said enzyme and determining the amount of said enzyme reaction product formed from a standard curve to provide a measure of the amount of collagenase inhibitor; and (d) comparing the amount of said collagenase inhibitor to that in serum or plasma from normal subjects, wherein an elevation of said amount of collagenase inhibitor is indicative of the presence of hepatic carcinoma. 12. The method of claim 11, wherein said solid substrate is a material selected from the group consisting of polystyrene, polycarbonate, polypropylene, and polyvinyl. 13. The method of claim 11, wherein said solid substrate is in a form selected from the group consisting of a ball, a microplate, a stick, and a test tube. 14. The method of claim 11, wherein said solid substrate is a polystyrene ball. 15. The method of claim 11, wherein said enzyme is horseradish peroxidase. 16. The method of claim 15, wherein said substrate is a substrate for horseradish peroxidase. 17. The method of claim 15, wherein said amount of said collagenase inhibitor is determined by measuring the optical density of the reaction mixture containing said enzyme reaction product formed at 450 nm. 18. The method of claim 11, wherein said standard curve is prepared by measuring the amount of enzyme reaction product formed in reaction mixtures containing varying amounts of human serum collagenase inhibitor. 19. The method of claim 11, wherein said screening further comprises performing liver function tests. 20. The method of claim 19, wherein said liver function tests are selected from the group consisting of the zinc sulfate turbidity test, measurement of glutamic oxaloacetic transaminase, measurement of glutamic-pyruvic transaminase, measurement of alkaline phosphatase, measurement of lactate dehydrogenase, measurement of .gamma.-glutamyl transpeptidase, measurement of prolyl hydroxylase, and measurement of .alpha.-fetoprotein.

Details for Patent 5,175,084

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2009-12-29