title: [Preparation,  identification and application  of   monoclonal antibody against matrix metalloproteinases-2].
abstract: OBJECTIVE
  To prepare and characterize monoclonal antibodies against matrix metalloproteinase-2 (MMP-2),  check its expression in the tissues of human ovarian cancer and transplanted tumors in nude mice.
  
  
  METHODS
  MMP-2 were linked to the carrier protein bovine serumalbumin (BSA) and keyhole limpet hemocyanin (KLH) using glutaraldehyde method to obtain MMP-2-BSA and MMP-2-KLH,  respectively. The anti-MMP-2 monoclonal antibody was obtained through hybridoma technique. We established the cell strains secreting mAb by hybridoma technique and prepared the mAb by induction of ascites in vivo. The prepared mAb was purified by salting out with ammonium sulfate and identified by ELISA and Western blotting. We compared the mAb and commercial polyclonal antibody by immunohistochemistry and detected the expressions of MMP-2 and CA125 in ovarian cancer issues and transplanted tumor.
  
  
  RESULTS
  The artificial antigen and 3 hybridoma cell lines secreting monoclonal antibodies (mAb) against MMP-2 were obtained. The subclasses of mAb were all  IgG1. The titer of peritoneal exudates was 1:1×10(6);. The expressions of MMP-2 and CA125 in transplanted tumor and ovarian cancer tissues were all high. The positive expression rate of MMP-2 checked using generated antibody was 71.2%(57/80) in ovarian cancer tissues and 16.67% (5/30) in normal tissues,  with significant difference between them (P<0.01). In early stage,  the positive rate of MMP-2 and CA125 combined detection was higher than that of CA125 detection alone (P<0.01). The mAb was suitable for detecting the expression of MMP-2 in human tissues and gave results consistent with commercial polyclonal antibody. The mAb was more specific than commercial mAb (P<0.01).
  
  
  CONCLUSIONS
  The anti-human MMP-2 mAb is successfully prepared,  which may serve as a valuable tool in the functionaI studies of ovarian cancer.
authors.affiliation: Department of Medicine, Huainan Union University, Huainan, China.
authors.forename: Mu
authors.initials: M
authors.lastname: You
authors.forename: Wen
authors.initials: W
authors.lastname: Liu
authors.forename: Zhen-shan
authors.initials: ZS
authors.lastname: Xu
authors.forename: Li-hua
authors.initials: LH
authors.lastname: Song
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D018394:CA-125 Antigen / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009368:Neoplasm Transplantation; D010051:Ovarian Neoplasms / Q000737:chemistry / Q000201:enzymology; D051381:Rats; D014183:Transplantation, Heterologous
chemical_list: D000911:Antibodies, Monoclonal; D018394:CA-125 Antigen; D020778:Matrix Metalloproteinase 2