Development of a monoclonal antibody-based immunochromatographic strip for cephalexin

Biomedical Chromatography

et al. 2015

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Rapid and sensitive indirect competitive enzyme-linked immunosorbent assays (ic-ELISA) and gold nanoparticle immunochromatographic strip tests were developed to detect thiamphenicol (TAP), florfenicol (FF) and chloramphenicol (CAP) in milk and honey samples. The generic monoclonal antibody for TAP, FF and CAP was prepared based on a hapten [D-threo-1-(4-aminophenyl)-2- dichloroacetylamino-1,3-propanediol], and the haptenwas linked to a carrier protein using the diazotization method. After the optimization of several parameters (coating, pH, sodium chloride content and methanol content), the ic-ELISA was established. The quantitative working range for TAP was 0.11-1.36 ng/mL, with an IC50 of 0.39 ng/mL. The optimized ELISA showed cross-reactivity to CAP (300%) and FF (15.6%), with IC50 values of 0.13 and 2.5 ng/mL, respectively. The analytical recovery of TAP, FF and CAP in milk and honey samples in the ic-ELISA ranged from 81.2 to 112.9%. Based on this monoclonal antibody, a rapid and sensitive immunochromatographic test strip was also developed. This strip had a detection limit of 1 ng/mL for TAP, FF and CAP in milk and honey samples. Moreover, the test was completed within 10 min. Our results showed that the proposed ic-ELISA and immunochromatographic test strip method are highly useful screening tools for TAP, FF and CAP detection in milk and honey samples.

“…The principle of the ICS is similar to the one-step competitive ELISA (Kuang et al, 2013;Guo et al, 2015). First, the sample solution and labeled-MAb were mixed and reacted for 5 min.…”

Section: Labeling Of the Mab With Colloidal Gold Particlesmentioning

confidence: 99%

Comparsion of an immunochromatographic strip with ELISA for simultaneous detection of thiamphenicol, florfenicol and chloramphenicol in food samples

Guo

Song

Biomedical Chromatography

et al. 2015

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“…18 In brief, CEX immunogen was prepared by conjugating CEX with KLH through DSS. 18 In brief, CEX immunogen was prepared by conjugating CEX with KLH through DSS.…”

Section: Preparation Of Anti-cex Monoclonal Antibodymentioning

confidence: 99%

Upconversion luminescence nanoparticles-based lateral flow immunochromatographic assay for cephalexin detection

Gao

et al. 2014

J. Mater. Chem. C

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Because of the near infrared excitation, upconversion luminescence (UCL) rare-earth nanoparticles are very suitable for biological applications in terms of low background interference. NaGdF 4 :Yb,Er nanoparticles were firstly synthesized through a replacement reaction at high temperature. To improve the upconversion luminescence efficiency, core-shell NaGdF 4 :Yb,Er@NaGdF 4 nanoparticles were then prepared by a seed-mediated method. The core-shell architecture significantly improved the upconversion luminescence and more effectively retained the luminescence during the following phase transfer process by ligand exchange. Upon further coupling with anti-cephalexin monoclonal antibody via "click" reaction, the resultant upconversion luminescence probe was obtained and used in a lateral flow immunochromatographic assay (LFIA) to detect the antibiotic residue of cephalexin. The results were compared with those achieved by a nanoprobe assay based on gold nanoparticles. More quantitative results were extracted by luminescent intensity analysis. Under optimized conditions, the detection limit of UCL nanoparticles-based LFIA was considerably comparable with gold nanoparticlesbased LFIA. † Electronic supplementary information (ESI) available: Shows the chemical stability and the optical stability of the conjugate of UCL particles with mAb. See

“…The ICA strip is suitable for qualitative detection, semiquantitative detection, and mass sample screenings. Results are visualized within 5-10 min by the naked eye (Dzantiev, Byzova, Urusov, & Zherdev, 2014;Guo et al, 2015;Sajid, Kawde, & Daud, 2015). Using a matched strip reader machine, the ICA strip becomes a quantification tool.…”

Section: Introductionmentioning

confidence: 99%

Development of ic-ELISA and lateral-flow immunochromatographic assay strip for the detection of folic acid in energy drinks and milk samples

Kong

Food and Agricultural Immunology

Song

et al. 2016

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Folic acid (FA) is an important vitamin for human growth and development, especially for pregnant women. A sensitive, rapid, and accurate FA detection method is required to assess the nutritional quality and safety of foods. A monoclonal antibody against FA was prepared for the development of indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and lateral-flow immunochromatographic assay (ICA) strip. The 50% inhibitory concentration and limit of detection of ic-ELISA were 0.12 and 0.018 ng/ml, respectively. The visual limit of detection and cut-off values of the lateral-flow ICA strip were 0.5 and 2.5 ng/ ml, respectively. Using the ICA strip, FA recovery rates were 89-98% from energy drinks and 73-87% for milk samples and were in good agreement with those obtained from the conventional microbiological assay method. Our developed methods are sensitive, convenient, effective, and suitable for on-site detection and rapid mass screening of food samples. ARTICLE HISTORY