Claims for Patent: 5,834,212

Summary for Patent: 5,834,212

Title:	Anti-human stromelysin monoclonal antibody and method for diagnosis of rheumatoid arthritis by enzyme immunoassay
Abstract:	Anti-human stromelysin monoclonal antibodies reactive with latent and active forms of stromelysin without discrimination between the two, each being immunoreactive with only one of the antigenic determinants of human stromelysin, are provided. The use of a combination of two such monoclonal antibodies which specifically react with different antigenic determinants of human stromelysin renders it possible to accurately determine the amount of human stromelysin in human body fluids, and thus to carry out the diagnosis of rheumatoid arthritis. There are thus provided said monoclonal antibodies per se, a sandwich enzyme immunoassay for the determination of the amount of human stromelysin in human body fluid samples using a combination of two such monoclonal antibodies, and a method for the diagnosis of rheumatoid arthritis using said immunoassay.
Inventor(s):	Okada; Yasunori (Matto-shi, Ishikawa-ken, 924, JP), Shinmei; Masayoshi (Tokorozawa-shi, Saitama-ken, 359, JP), Hayakawa; Taro (Tenpaku-ku, Nagoya-shi, Aichi-ken, 468, JP), Iwata; Kazushi (Takaoka-shi, Toyama-ken, 933, JP), Korin; Yumi (Kahoku-gun, Ishikawa-ken, 929-12, JP), Kodama; Shuji (Takaoka-shi, Toyama-ken, 933, JP), Yoshida; Shinichi (Toyama-shi, Toyama-ken, 930, JP)
Assignee:
Application Number:	08/417,847
Patent Claims:	1. A monoclonal antibody which specifically binds to latent stromelysin (prostromelysin) and active stromelysin without discrimination between the two, but which does not bind to either collagenase or gelatinase, wherein said monoclonal antibody is produced by a hybridoma which is selected from the group consisting of hybridoma 55-2A4, FERM BP-3743, and hybridoma 55-3G3, FERM BP-3744. 2. The monoclonal antibody according to claim 1, wherein said hybridoma is hybridoma 55-2A4, FERM BP-3743. 3. The monoclonal antibody according to claim 1, wherein said hybridoma is hybridoma 55-3G3, FERM BP-3744. 4. A sandwich enzyme immunoassay for determining the amount of active stromelysin or prostromelysin in a sample selected from the group consisting of blood, serum and synovial fluid, comprising the steps of: (a) contacting the sample suspected of containing active stromelysin or prostromelysin with an enzyme-labeled monoclonal antibody or Fab' fraction thereof, wherein said monoclonal antibody or Fab' fraction thereof specifically binds to a first region of active stromelysin or prostromelysin without discrimination between the two, but does not bind to either collagenase or gelatinase, to form complexes of said active stromelysin or said prostromelysin and said monoclonal antibody or said Fab' fraction thereof; (b) contacting said complexes of step (a) with an immobilized monoclonal antibody which specifically binds to a second region of said active stromelysin or prostromelysin without discrimination between the two, and which also does not bind to either collagenase or gelatinase, so as to form immobilized complexes of said enzyme-labeled monoclonal antibody or Fab' fraction thereof, said active stromelysin and/or said prostromelysin, and said immobilized monoclonal antibody; (c) washing said complexes of step (b); (d) incubating said complexes of step (c) with a substrate of said enzyme to from an amount of end product; (e) determining the amount of end product formed in step (d); and (f) relating said amount of end product determined in step (e) as indicative of the amount of active stromelysin and prostromelysin in said sample by referring to a standard curve for prostromelysin, wherein said enzyme-labeled monoclonal antibody is a monoclonal antibody produced by hybridoma 55-2A4, FERM BP-3743, and wherein said immobilized monoclonal antibody is a monoclonal antibody produced by hybridoma 55-3G3, FERM BP-3744. 5. The sandwich enzyme immunoassay of claim 4, wherein said immobilized monoclonal antibody is immobilized on a solid support selected from the group consisting of a sphere, a microplate, a stick, and a test tube. 6. The sandwich enzyme immunoassay of claim 5, wherein said solid support is made of a material selected from the group consisting of a polystyrene, a polycarbonate, a polypropylene, and a polyvinyl. 7. The sandwich enzyme immunoassay of claim 4, wherein said enzyme-labeled monoclonal antibody or Fab' fraction thereof is labeled with an enzyme selected from the group consisting of peroxidase, alkaline phosphatase, and .beta.-D-galactosidase. 8. The sandwich enzyme immunoassay of claim 4, wherein said determining of step (e) is performed by colorimetry, fluorometry, bioluminescence, or chemiluminescence. 9. A method for diagnosing rheumatoid arthritis in a subject, comprising determining the amount of active stromelysin and prostromelysin in a sample of blood, serum or synovial fluid obtained from the subject by said sandwich enzyme immunoassay of claim 4, and comparing said amount with that determined in normal subjects, wherein an elevated amount of active stromelysin and/or prostromelysin in the sample is indicative of rheumatoid arthritis in said subject.

Details for Patent 5,834,212

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2015-11-10