title: Establishment and characterization of 7 new monoclonal antibodies to tissue inhibitor of metalloproteinases-1.
abstract: Tissue inhibitor of metalloproteinases-1 (TIMP-1) plays a pivotal role in tissue remodeling processes, such as inflammation, wound healing and cancer invasion. Experimental results have pointed to a role in angiogenesis, cell proliferation, apoptosis and in malignant transformation. In clinical investigations high tumor tissue or plasma levels of TIMP-1 have been shown to have a strong and independent association with shorter survival time for breast and colorectal cancer patients, respectively. The purpose of this study has been to develop and characterize new anti-TIMP-1 monoclonal antibodies that may be useful in future development of TIMP-1 immunoassays.Peptide-based epitope mapping reveals linear epitopes. Surface plasmon resonance was used to determine antibody affinity and ability of antibodies to sandwich with each other. Antigen recognition was tested using ELISA and a chemiluminescence microtiter immunoassay format. Three antibodies recognized linear peptides. Estimated antibody affinities for TIMP-1 ranged from 6.6 x 10(8) to>10(10) 1/M. Antibodies demonstrated different abilities in 'capture' and 'detection' positions in the sandwich experiment. All antibody pairs bound TIMP-1:ProMMP-9 complexes. TIMP-1:MMP-9 complexes were marginally reactive with five antibody pairs. The results suggest that the antibodies are unique. They may be useful in designing assays that recognize various forms of TIMP-1. Future studies will clarify whether the use of different combinations of antibodies will increase the clinical value of TIMP-1 measurements in the treatment of cancer patients.
authors.affiliation: Department of Veterinary Pathobiology, Royal Veterinary and Agricultural University, Frederiksberg C, Denmark. nas@kvl.dk
authors.forename: Nanna
authors.initials: N
authors.lastname: Møller Sørensen
authors.forename: Barry L
authors.initials: BL
authors.lastname: Dowell
authors.forename: Kent D
authors.initials: KD
authors.lastname: Stewart
authors.forename: Vibeke
authors.initials: V
authors.lastname: Jensen
authors.forename: Lise
authors.initials: L
authors.lastname: Larsen
authors.forename: Ulrik
authors.initials: U
authors.lastname: Lademann
authors.forename: Gillian
authors.initials: G
authors.lastname: Murphy
authors.forename: Hans Jørgen
authors.initials: HJ
authors.lastname: Nielsen
authors.forename: Nils
authors.initials: N
authors.lastname: Brünner
authors.forename: Gerard J
authors.initials: GJ
authors.lastname: Davis
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006801:Humans; D006825:Hybridomas; D008163:Luminescent Measurements; D020780:Matrix Metalloproteinase 9 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation; D020349:Surface Plasmon Resonance; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D019715:Tissue Inhibitor of Metalloproteinase-1; D020780:Matrix Metalloproteinase 9