title: Production and characterization of murine monoclonal anti-human DNase II antibodies, and their use for immunoaffinity purification of DNase II from human liver and urine.
abstract: Four murine monoclonal anti-human deoxyribonuclease II (DNase II) antibodies were obtained from BALB/c mice immunized with human DNase II purified from human liver. Both single radial enzyme diffusion (SRED) and DNA-cast polyacrylamide gel electrophoresis (DNA-cast PAGE) were very useful for obtaining the DNase II-specific antibodies. All of the antibodies showed specific inhibition of human DNase II enzyme activity and specific immunostaining of the 32-kDa enzyme band, which is one of the three non-identical subunits of human DNase II molecule separated by sodium dodecyl sulfate (SDS)-PAGE followed by blotting on a transfer membrane. A formyl-cellulofine resin conjugated with each antibody specifically adsorbed and efficiently desorbed the active DNase II enzyme. Insertion of the immunoaffinity step in our purification procedure made the purification of human DNase II easier, faster and more effective than the conventional procedure.
authors.affiliation: Department of Legal Medicine, Gunma University School of Medicine, Maebashi, Gunma 371-8511, Japan.
authors.forename: Tamiko
authors.initials: T
authors.lastname: Nakajima
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Yasuda
authors.forename: Haruo
authors.initials: H
authors.lastname: Takeshita
authors.forename: Shinjiro
authors.initials: S
authors.lastname: Mori
authors.forename: Kouichi
authors.initials: K
authors.lastname: Mogi
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kaneko
authors.forename: Emiko
authors.initials: E
authors.lastname: Nakazato
authors.forename: Koichiro
authors.initials: K
authors.lastname: Kishi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000737:chemistry / Q000276:immunology*; D005260:Female; D006801:Humans; D008099:Liver / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D004706:Endodeoxyribonucleases; C019329:deoxyribonuclease II