Synthesis of Haptens and Development of an Immunoassay for the Olive Fruit Fly Pheromone
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- 作者：Afroditi Neokosmidi ; Valentine Ragoussis ; Christos Zikos ; Maria Paravatou-Petsotas ; Evangelia Livaniou ; Nikitas Ragoussis ; and Gregory Evangelatos
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2004
- 出版时间：July 14, 2004
- 年：2004
- 卷：52
- 期：14
- 页码：4368 - 4374
- 全文大小：99K
- 年卷期：v.52,no.14(July 14, 2004)
- ISSN：1520-5118
文摘
An enzyme-linked immunosorbent assay (ELISA) for the olive fruit fly pheromone, Bactrocera oleaeGmelin, was developed. The assay uses polyclonal antibodies, raised in rabbits, against (±)-
-[3-(1,7-dioxaspiro[5.5]undecane)]propionic acid, 2 (hapten I), conjugated to the KLH (keyhole limpethemocyanin) by the carbodiimide method. A second hapten, (±)-
-[3-(1,7-dioxaspiro[5.5]undecane)]butylamine, 3 (hapten II), after conjugation to a biotin moiety, was used for indirect immobilizationonto ELISA microwells precoated with the glycoprotein avidin. The developed ELISA method measuresthe synthetic olive fruit fly pheromone in concentrations ranging between 0.08 and 10 
g/mL andshows great promise for practical applications for pheromone detection in environmental and biologicalsamples. The results obtained strongly indicate that this technique, to our knowledge the first insectpheromone enzyme-linked immunosorbent assay so far reported, is a fast, sensitive, inexpensive,and highly convenient method for the analysis of a volatile and low molecular weight compound suchas 1,7-dioxaspiro[5.5]undecane, 1.Keywords: Olive fruit fly; Bactrocera oleae (Gmelin); pheromones; hapten; antipheromone polyclonalantibodies; enzyme-linked immunosorbent assay (ELISA); avidin-biotin system
-[3-(1,7-dioxaspiro[5.5]undecane)]propionic acid, 2 (hapten I), conjugated to the KLH (keyhole limpethemocyanin) by the carbodiimide method. A second hapten, (±)--[3-(1,7-dioxaspiro[5.5]undecane)]butylamine, 3 (hapten II), after conjugation to a biotin moiety, was used for indirect immobilizationonto ELISA microwells precoated with the glycoprotein avidin. The developed ELISA method measuresthe synthetic olive fruit fly pheromone in concentrations ranging between 0.08 and 10 g/mL andshows great promise for practical applications for pheromone detection in environmental and biologicalsamples. The results obtained strongly indicate that this technique, to our knowledge the first insectpheromone enzyme-linked immunosorbent assay so far reported, is a fast, sensitive, inexpensive,and highly convenient method for the analysis of a volatile and low molecular weight compound suchas 1,7-dioxaspiro[5.5]undecane, 1.Keywords: Olive fruit fly; Bactrocera oleae (Gmelin); pheromones; hapten; antipheromone polyclonalantibodies; enzyme-linked immunosorbent assay (ELISA); avidin-biotin system