Abstract
ABSTRACT Immunoassay direct for 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ), the metabolite of furaltadone, based on a specific antibody was usually unavailable due to its small molecule weight. In this work, a hapten with a double bond and an active carboxyl group was designed and then conjugated to peptide dendrimer to obtain novel multiple haptens. The haptens and multiple haptens were coupled to bovine serum albumin as immunogens. Polyclonal antibodies showed binding to free AMOZ in a heterologous competitive indirect enzyme-linked immunosorbent assay were obtained. The assay showed IC50 (half maximal inhibitory concentration) of 4.1 μg/kg and limit of detection of 0.2 μg/kg for AMOZ. Recoveries of AMOZ from grass carp were tested from 83.1% to 117.0%, with the coefficient of variation below 12%. As no laborious derivatisation procedure was employed, the proposed assay should be useful for the screening of AMOZ for a large number of samples.
Highlights
Immunoassay based on antigen–antibody interaction has many excellent features, such as being accurate, quick, sensitive and high throughput (Gomaa & Boye, 2015; Kong et al, 2017; Shen et al, 2017; Tighe, Ryder, Todd, & Fairclough, 2015)
For AMOZ, a derivative strategy was adopted in most immunoassays (Pimpitak et al, 2009; Shen et al, 2012)
Due to the unique molecular characteristics of dendrimers, they demonstrated the ability to enhance the immunogenicity of vaccine antigen for immunoassay
Summary
Immunoassay based on antigen–antibody interaction has many excellent features, such as being accurate, quick, sensitive and high throughput (Gomaa & Boye, 2015; Kong et al, 2017; Shen et al, 2017; Tighe, Ryder, Todd, & Fairclough, 2015). It has been successfully applied in detecting low molecular mass organic contaminations in food, especially in screening large numbers of samples, playing an important role in food safety regulation (Anfossi, Di Nardo, Giovannoli, Passini, & Baggiani, 2015; Bolarinwa, Orfila, & Morgan, 2014; Gujral, Yoo, Bamdad, Suh, & Sunwoo, 2017; Xie, Kong, Liu, Song, & Kuang, 2017). Due to the lack of epitope of haptens, the production of antibodies with high affinity and high specificity is still one of the bottlenecks of immunoassay for some small molecules
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