Use of human recombinant DNase I expressed in COS-7 cells as an immunogen to produce a specific anti-DNase I antibody.
To obtain human deoxyribonuclease I (DNase I) as an immunogen, we have developed a procedure that is more useful and effective than the conventional procedure, which uses human urine as a starting material. In the new procedure, we culture COS-7 cells transfected with expression vector carrying human DNase I cDNA, and then purify the enzyme from the culture medium. The enzyme can be easily isolated to apparent homogeneity by passage through only three chromatography columns. The rabbit antiserum that we used against the recombinant DNase I was not inferior to that used against DNase I from human urine, in terms of both its ability to discriminate DNase I phenotypes and its ability to neutralize enzyme activity. Therefore, our procedure may be useful for producing an antibody specific for human DNase I.
Takeshita
H
H
Department of Legal Medicine, Gunma University School of Medicine, Gunma, Japan.
Yasuda
T
T
Nakazato
E
E
Nakajima
T
T
Mori
S
S
Mogi
K
K
Kaneko
Y
Y
Iida
R
R
Kishi
K
K
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D000941:Antigens / Q000235:genetics; D001483:Base Sequence; D019556:COS Cells; D018076:DNA, Complementary / Q000235:genetics; D003850:Deoxyribonuclease I / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000652:urine; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D007114:Immunization; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D000941:Antigens; D018076:DNA, Complementary; D011994:Recombinant Proteins; D003850:Deoxyribonuclease I