CONSTRUCTION OF SANDWICH ENZYME-IMMUNOASSAY FOR RAT TRANSTHYRETIN USING RECOMBINANT ANTIGEN APPROACH, AND ITS APPLICATION

被引：4

作者：

MATSUMOTO, T

MORITA, M

SHIRAI, H

KISHI, T

机构：

[1] Research Laboratory of Applied Biochemistry, Tanabe Seiyaku Co. Ltd., Osaka 532, 16-89 Kashima 3-chome, Yodogawa-ku

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 1993年 / 57卷 / 03期

关键词：

D O I：

10.1271/bbb.57.414

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：

A sandwich enzyme immunoassay for rat transthyretin using a recombinant antigen approach is described. Rat transthyretin cDNA was cloned by PCR. Rat transthyretin fused with the IgG-binding ZZ domain of protein A was expressed in E. coli using the fusion vector pEZZ18. Antibody against the fusion protein was specific to rat transthyretin in plasma as shown by immunoblotting. Affinity-purified anti-rat transthyretin-ZZ was biotinylated and used for the sandwich enzyme immunoassay. In this assay, the measurable range was 1.2-50 ng/ml and the coefficients of variation within and between the assay series (assay range: 2.5-20ng/ml) were 1.31+/-0.08% and 3.50+/-1.46%, respectively. Cross-reactivity was examined using bovine, human, and mouse serum. There was a cross-reaction only with mouse transthyretin. In an in vitro experiment, transthyretin secreted by rat hepatocytes could be measured by the sandwich enzyme immunoassay.

引用

页码：414 / 418

页数：5

共 21 条

[11]

MILANO G, 1978, J NATL CANCER I, V61, P687

[12] CLONING AND SEQUENCE-ANALYSIS OF CDNA FOR HUMAN PREALBUMIN [J].

MITA, S ;

MAEDA, S ;

SHIMADA, K ;

ARAKI, S .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1984, 124 (02) :558-564

[13]

NAVAB M, 1977, J BIOL CHEM, V252, P5100

[14]

OPPENHEI.JH, 1965, J BIOL CHEM, V240, P173

[15]

Peterson P A, 1974, Vitam Horm, V32, P181

[16]

PETERSON PA, 1973, J BIOL CHEM, V248, P4009

[17]

SARAIVA MJM, 1987, NEUROLOGY, V48, P189

[18]

SCHREIBER G, 1987, PLASMA PROTEINS STRU, V5, P293

[19]

SUNDELIN J, 1985, J BIOL CHEM, V260, P6841

[20] ELECTROPHORETIC TRANSFER OF PROTEINS FROM POLYACRYLAMIDE GELS TO NITROCELLULOSE SHEETS - PROCEDURE AND SOME APPLICATIONS [J].

TOWBIN, H ;

STAEHELIN, T ;

GORDON, J .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (09) :4350-4354

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