Development of a neutralizing antibody specific for the active form of matrix metalloproteinase-13.
Matrix metalloproteinase-13 (MMP-13) is important in the pathology of osteoarthritis (OA). Although MMP-13 is considered a therapeutic target for OA, it is unclear how MMP-13 activity is regulated by the system that comprises various proteinases and their inhibitors. MMP-13 neutralizing antibodies could be a useful tool for investigating the involvement of MMP-13 in cartilage degeneration in OA-affected joints because antibodies possess high affinity and specificity compared with low-molecular weight chemical compounds. On the basis of three-dimensional structure and amino acid sequence information on MMP-13, we selected an appropriate antigen peptide and generated a neutralizing antibody by immunizing mice with the antigen. The most significant property of monoclonal antibody 14D10 was the specific binding to the active form of MMP-13, but not to the latent form, or other MMPs. With this property, active MMP-13 was measured selectively by an enzyme-linked immunosorbet assay. Furthermore, 14D10 suppressed the cleavage of type II collagen in human articular chondrocyte cultures, and 14D10 is thought to inhibit MMP-13 activity effectively. Thus, the highly selective MMP-13 neutralizing antibody (14D10) might be a useful tool for investigating the mechanism of type II collagen degradation in arthritic pathology.
Shionogi Pharmaceutical Research Center, Shionogi & Company, Ltd., 3-1-1 Futaba-cho, Toyonaka, Osaka 561-0825, Japan.
Shoichi
S
Naito
Tatsuya
T
Takahashi
Junji
J
Onoda
Akira
A
Yamauchi
Taeko
T
Kawai
Junji
J
Kishino
Shoji
S
Yamane
Ikuo
I
Fujii
Naoshi
N
Fukui
Yoshito
Y
Numata
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D057134:Antibodies, Neutralizing; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism; D019902:Chondrocytes / Q000378:metabolism; D024043:Collagen Type II / Q000378:metabolism; D005260:Female; D006801:Humans; D053509:Matrix Metalloproteinase 13 / Q000276:immunology*; D051379:Mice; D010003:Osteoarthritis / Q000378:metabolism
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D024043:Collagen Type II; D053509:Matrix Metalloproteinase 13