BIOLUMINESCENT IMMUNOASSAY USING A MONOMERIC FAB'-PHOTOPROTEIN AEQUORIN CONJUGATE

被引：23

作者：

ERIKAKU, T

ZENNO, S

INOUYE, S

机构：

[1] Yokohama Research Center, Chisso Corporation, Kanazawa-ku, Yokohama, 236

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1991年 / 174卷 / 03期

关键词：

D O I：

10.1016/0006-291X(91)91568-W

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The photoprotein aequorin emits light by an intermolecular reaction when mixed with Ca2+. To apply a bioluminescent immunoassay based on the light emission property of aequorin, we prepared aequorin-antibody Fab' conjugates by a chemical cross-linking technique. Recombinant apoaequorin was coupled with human tumor necrosis factor-α (TNF-α) antibodies (polyclonal Fab' or monoclonal Fab' fragments) using N-succinimidyl 4-(N-maleimidemethyl)cyclohexane-1-carboxylate. The luminescent activity of the Fab'-aequorin conjugate was about one-tenth that of aequorin. Using a monomeric conjugate, human TNF-α can be measured at an attomole level by a sandwich immunoassay technique. © 1991.

引用

页码：1331 / 1336

页数：6

共 21 条

[1]

BLINKS JR, 1976, PHARMACOL REV, V28, P1

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3] EXPRESSION AND SECRETION OF AEQUORIN AS A CHIMERIC ANTIBODY BY MEANS OF A MAMMALIAN EXPRESSION VECTOR [J].