PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST HUMAN CU/ZN SUPEROXIDE-DISMUTASE AND THE ESTABLISHMENT OF A SUPER-RAPID ENZYME-LINKED-IMMUNOSORBENT-ASSAY (SURALISA)

被引:17
作者
PORSTMANN, T
WIETSCHKE, R
GRUNOW, R
JAHN, S
PORSTMANN, B
SCHMECHTA, H
PERGANDE, M
VONBAEHR, R
机构
[1] HUMBOLDT UNIV,BEREICH MED CHARITE,SCH MED,DEPT PATHOL & CLIN BIOCHEM,O-1040 BERLIN,GERMANY
[2] HUMBOLDT UNIV,BEREICH MED CHARITE,DEPT EXPTL ORGAN TRANSPLANTAT,O-1040 BERLIN,GERMANY
[3] MIL MED ACAD,INST FORENS MED,BAD SAAROW,GERMANY
关键词
Cu/Zn superoxide dismutase; human; recombinant; ELISA; additive; super-rapid; two-site; Epitope analysis; Monoclonal antibody; Polyclonal antibody;
D O I
10.1016/0022-1759(90)90333-Q
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Murine monoclonal IgG1 antibodies directed against four different epitopes of human Cu/Zn superoxide dismutase (SOD) were produced by immunization with recombinant Cu/Zn SOD. The antibodies reacted well with the recombinant protein and Cu/Zn SOD purified from human erythrocytes, with binding constants ranging from 8.8 × 109 to 2.2 × 1010 1/mol. When mixed, these antibodies completely prevented the binding of rabbit and sheep polyclonal antibodies raised against erythrocyte Cu/Zn SOD. Whereas one antibody was directed against a common homology region of bovine and human Cu/Zn SOD, all the other antibodies reacted exclusively or preferentially with human Cu/Zn SOD. Only one epitope on the human Cu/Zn SOD molecule was accessible at two different sites as demonstrated in a homologous two-site assay with one and the same antibody used as both capture and indicator antibody. In the indirect two-site assay with unlabelled monoclonal antibodies, an additive effect with a steeper dose-response curve was obtained by mixing antibodies against different epitopes. A super-rapid one-step two-site enzyme immunoassay (overall duration 20 min) was established with antibodies against two different epitopes. Its detection limit was 0.5 μg SOD/1. © 1990.
引用
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页码:1 / 10
页数:10
相关论文
共 25 条
[21]  
PORSTMANN T, 1985, J CLIN CHEM CLIN BIO, V23, P41
[22]  
RIDROVICH I, 1974, OXYGEN LIVING PROCES, P250
[23]   TRISOMY-21 AND SUPEROXIDE DISMUTASE-1 (IPO-A) - ATTEMPT AT LOCALIZATION ON SUB-BAND 21Q 22.1 [J].
SINET, PM ;
COUTURIER, J ;
DUTRILLAUX, B ;
POISSONNIER, M ;
RAOUL, O ;
RETHORE, MO ;
ALLARD, D ;
LEJEUNE, J ;
JEROME, H .
EXPERIMENTAL CELL RESEARCH, 1976, 97 (01) :47-55
[24]  
STANSELL MJ, 1965, J BIOL CHEM, V240, P4299
[25]   LINKAGE OF GENES FOR HUMAN INTERFERON-INDUCED ANTIVIRAL PROTEIN AND INDOPHENOL OXIDASE-B TRAITS TO CHROMOSOME G-21 [J].
TAN, YH ;
TISCHFIE.J ;
RUDDLE, FH .
JOURNAL OF EXPERIMENTAL MEDICINE, 1973, 137 (02) :317-330