Detection of heterogeneity of Cu, Zn‐superoxide dismutase with monoclonal antibodies and the establishment of a highly sensitive fluorescence sandwich enzyme‐linked immunosorbent assay

FEBS Letters

Ono, Kumeo¹  Naruse, Takuji¹  Nakano, Minoru²  Kimura, Satsuki² 

[1] The Third Department of Internal Medicine, Gunma, University, Maebashi, Gunma, Japan;College of Medical Care and Technology, Gunma University, Maebashi, Gunma, Japan

关键词: Cu;    Zn-superoxide dismutase;    Monoclonal antibody;    Immunoassay;    SOD;    superoxide dismutase;    Ab;    antibody;    mAb;    monoclonal antibody;    MCLA;    2-methyl-6-(p-methoxyphenyl)-3;    7-dihydroimidazo[1;    2-α]pyrazin-3-one;    ELIZA;    enzyme linked immunosorbent assay;   

DOI  :  10.1016/0014-5793(91)80457-E

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Murine monoclonal antibodies directed against a native form of Cu, Zn-superoxide dismutase (SOD) were produced by immunizing SOD purified from human erythrocytes. The monoclonal antibodies able to bind SOD were further screened for their ability to absorb SOD activity using antimouse 1gG conjugated iron beads as solid supports in magnetic separation. This new screening method revealed the heterogeneity of native SOD in the reactivity with the antibodies. One monoclonal antibody succesfully absorbed the entire activity of SOD detected by an inhibition assay of cypridina luciferin analog (MCLA)-dependent chemilluminescence induced by superoxide anion production, while others absorbed only a part of the SOD activity. The evidence that all of the latter antibodies failed to react with recombinant artificial SOD free of charge isomers suggested correlation of the heterogeneity with the presence of charge isomeric forms. The former antibody was further used to establish a fluorescence sandwich enzyme immunoassay, and this provided a very sensitive detection limit as low as 100 pg/ml.

【 授权许可】

Unknown