PURPOSE: To obtain a new monoclonal antibody obtained in the presence of a bone sialoprotein purified from bovine bone as an immunogen, recognizing a characteristic bone protein existing in human blood or its metabolites, and useful for diagnosing the composition changes of bones, the abnormal metabolisms of bone tissues, etc.
CONSTITUTION: This monoclonal antibody is obtained by grinding bovine bone in liquid nitrogen, washing the obtained bone powder, defatting the washed bone powder, deashing the defatted bone powder, extracting the obtained deashed bone substrate with a buffer solution containing 4M guanidine hydrochloride, successively feeding the extract into a hydroxyapatite column, a heparin- immobilized column, a gel filtration column and a reverse phase high performance chromatography, immunizing a mouse with the purified bone sialoprotein, collecting the splenic cells of the immunized mouse, fusing the collected splenic cells with myeloma cells, selectively culturing the fused cells in a HAT medium, screening an antibody-producing strain, cloning the screened cell, and subsequently culturing the cloned cell to obtain the objective monoclonal antibody recognizing the characteristic bone protein existing in human blood or its metabolites.
MATSUYAMA TOSHIKATSU
TAIRA TOSHIO
