Preparation Of Canine Matrix Metalloproteinase 13 Monoclonal Antibody And Study Of Osteochondrosis

Osteochondrosis(OC)is a disease that severely affects the movement of animals.The diseased animals eventually show clinical manifestations with different degrees of severity characterized by claudication,motor ataxia and joint deformation.At present,the pathogenesis of osteochondrosis is not clear,and it is generally believed that osteochondrosis is caused by multiple factors.Dog osteochondral cartilage is composed of chondrocytes and ECM.The main components of ECM are collagen type II and glycosaminoglycan.MMP-13 may play an important role in the occurrence and development of canine osteochondrosis.However,there is currently no monoclonal antibody for detection of MMP-13 in dogs.The purpose of this study is to obtain anti-MMP-13 monoclonal antibody to explore the pathogenesis of canine osteochondrosis.This study for dog MMP-13 genes of CDS,using prokaryotic expression system were successfully induced and MMP-13 recombinant protein.using recombinant proteins immuned mice,won the 10 strains by the clone selection could stable secret anti-MMP-13 monoclonal antibody hybridoma cell lines,the antibody titer of 819200,subtypes of Ig G1.Immunohistochemical staining results showed that the expression of MMP-13 was mainly distributed in the surface and middle chondrocytes cytoplasm and cartilage lacunae area near the injury site.Western blotting assay showed that MMP-13 increased significantly on day 30 and day 60 compared with the sham group.This study of MMP-13 monoclonal antibody was successfully achieved and verified in the model of osteochondrosis in the dog,and reveals the model of osteochondrosis of MMP-13 distribution in the model and relative content in serum of the schedule change,for early diagnosis and treatment of osteochondrosis,provides a theoretical basis to take prevention measures in advance,slow or reverse the disease process,reduces the economic loss,improve the animal welfare.