Comparative sensitivity of immunoassays for haptens using monomeric and dimeric antibody fragments

被引：22

作者：

Grant, SD ^{[1
]}

Porter, AJ ^{[1
]}

Harris, WJ ^{[1
]}

机构：

[1] Univ Aberdeen, Inst Med Sci, Dept Mol & Cell Biol, Aberdeen AB25 2ZD, Scotland

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 1999年 / 47卷 / 01期

关键词：

antibody fragments; immunoassays; single-chain antibodies; anti-haptens; triazines;

D O I：

10.1021/jf9808574

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

A single-chain anti-atrazine antibody fragment, scAb (single-chain Fv with a CK domain), was expressed in Escherichia coli, and monomeric and dimeric species were preferentially purified from periplasmic extracts by chromatography upon nickel chelate immunosorbent columns or by immunoaffinity purification using a constant domain (CK) tag. Recombinant monomeric and dimeric antibody fragments, Fab, and intact monoclonal antibodies were compared in assays by competition between free atrazine in solution and (a) immobilized atrazine-bovine serum albumin conjugate (indirect assay) or (b) atrazine-alkaline phosphatase (direct assay). Recombinant antibody fragments provided a lower detection Limit than either Fab or intact monoclonal antibody in both assay formats. Monomeric fragments displayed a sensitivity of detection down to 0.1 ppb, compared to 1.0 ppb for dimeric fragments and the parental monoclonal.

引用

页码：340 / 345

页数：6

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