Rapid purification of human DNase I using mouse monoclonal anti-DNase I antibodies and characterization of the antibodies |
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Authors: | Nakajima T Yasuda T Takeshita H Nakashima Y Mori S Mogi K Kishi K |
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Affiliation: | Department of Legal Medicine, Gunma University School of Medicine, Maebashi, Japan. |
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Abstract: | Five anti-human deoxyribonuclease I (DNase I) monoclonal antibodies were obtained from BALB/c mice immunized with DNase I purified from human urine. Four of them inhibited DNase I enzyme activity, as did a rabbit polyclonal antibody; these 4 did not have immunostaining ability. The remaining one had immunostaining ability but no inhibitory activity. A Sepharose 4B column conjugated with 1 of the 4 antibodies that had inhibitory activity effectively adsorbed and eluted the DNase I enzyme; this did not occur with the rabbit polyclonal antibody. We showed that adding an immunoaffinity chromatography step made the purification of human DNase I easier and faster than the conventional procedure. |
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