[Production of monoclonal antibody to chicken epiphyseal cartilage matrix vesicle metalloprotease to determine its localization].
Previous reports demonstrated that metalloprotease was isolated from matrix vesicles (MVs) of chicken epiphyseal cartilage. In this study, we produced the monoclonal antibody against MV metalloprotease (MVPase) and its location was studied by immunohistochemical technique. Isolated MVs were injected into Balb/c mice and spleen cells were collected for fusion with myeloma cells. A monoclonal hybrid cell producing antibody against MVPase was isolated. The immunoglobulin isotype was IgG1. On western blots, this antibody reacted with a single band protein of 33KDa which corresponded with purified MVPase. In the light immunohistochemical study, immuno reactions were observed in the mesenchymal cells of bone marrow at the lowest area in hypertrophic zone of the epiphyseal cartilage. Ultrastructural immunolocalization of protein A-gold particles revealing MVPase showed that MVs contained this enzyme, and in the calcifying area, gold particles were observed at the apatite needles of ruptured MVs. According to these observations, it is suggested that MVPase was already synthesized in undifferentiated marrow bone cells and this enzyme promotes the initial MV calcification and relates closely to the growth crystalline needles of apatite.
Sadamatsu
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Department of Orthopaedic Surgery, Nagasaki University School of Medicine, Japan.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002645:Chickens; D006132:Growth Plate / Q000201:enzymology / Q000276:immunology*; D008666:Metalloendopeptidases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D008666:Metalloendopeptidases