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
收藏
页码：340 / 345
页数：6
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