Rapid purification of human DNase I using mouse monoclonal anti-DNase I antibodies and characterization of the antibodies.
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.
Nakajima
T
T
Department of Legal Medicine, Gunma University School of Medicine, Maebashi, Japan.
Yasuda
T
T
Takeshita
H
H
Nakashima
Y
Y
Mori
S
S
Mogi
K
K
Kishi
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002846:Chromatography, Affinity / Q000379:methods; D003850:Deoxyribonuclease I / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D013997:Time Factors
D000911:Antibodies, Monoclonal; D003850:Deoxyribonuclease I