Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Pentachloronitrobenzene Residues in Environmental Samples

Xu, Ting; Shao, Xiao Ling; Li, Qing X.; Keum, Young Soo; Jing, Hong; Sheng, Wei; Li, Ji

doi:10.1021/jf063287d

Cited by 10 publications

(7 citation statements)

References 20 publications

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“…Accordingly, we studied organic solvent, pH, ionic strength, detergent, and blocking reagent as possible sources of interference in the ELISA with DBP MAb. Most researchers reached the same conclusion that methanol caused the least negative effect on the ELISA performance [33]–[35]. However, our assays showed that DMF presented the best combination of A max and IC 50 .…”

Section: Discussionsupporting

confidence: 70%

“…The ELISA for DBP was very sensitive to ionic strength, that is, A max and IC 50 values significantly decreased at high concentration of NaCl. The sensitivity improvement by increased ionic strength may be due to dispersion and weakening of the nonspecific binding derived from antibody [35]. Tween-20 is a nonionic detergent commonly used in washing buffer to reduce nonspecific interactions in ELISA.…”

Section: Discussionmentioning

confidence: 99%

“…Generally, the sensitivity and CR of antibody are dependent on several factors such as the structure of immunogen and the physicochemical properties of the test analytes. Electrostatic and hydrophobic interaction can determine the strength of binding and CR through the difference of electron distribution between haptens [35]. MAb was raised against DBAP-OVA conjugate in this study.…”

Section: Discussionmentioning

confidence: 99%

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An Immunoassay for Dibutyl Phthalate Based on Direct Hapten Linkage to the Polystyrene Surface of Microtiter Plates

et al. 2011

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BackgroundDibutyl phthalate (DBP) is predominantly used as a plasticizer inplastics to make them flexible. Extensive use of phthalates in both industrial processes and other consumer products has resulted in the ubiquitous presence of phthalates in the environment. In order to better determine the level of pollution in the environment and evaluate the potential adverse effects of exposure to DBP, immunoassay for DBP was developed.Methodology/Principal FindingsA monoclonal antibody specific to DBP was produced from a stable hybridoma cell line generated by lymphocyte hybridoma technique. An indirect competitive enzyme-linked immunosorbent assay (icELISA) employing direct coating of hapten on polystyrene microtiter plates was established for the detection of DBP. Polystyrene surface was first oxidized by permanganate in dilute sulfuric acid to generate carboxyl groups. Then dibutyl 4-aminophthalate, which is an analogue of DBP, was covalently linked to the carboxyl groups of polystyrene surface with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). Compared with conjugate coated format (IC50 = 106 ng/mL), the direct hapten coated format (IC50 = 14.6 ng/mL) improved assay sensitivity after careful optimization of assay conditions. The average recovery of DBP from spiked water sample was 104.4% and the average coefficient of variation was 9.95%. Good agreement of the results obtained by the hapten coated icELISA and gas chromatography-mass spectrometry further confirmed the reliability and accuracy of the icELISA for the detection of DBP in certain plastic and cosmetic samples.Conclusions/SignificanceThe stable and efficient hybridoma cell line obtained is an unlimited source of sensitive and specific antibody to DBP. The hapten coated format is proposed as generally applicable because the carboxyl groups on modified microtiter plate surface enables stable immobilization of aminated or hydroxylated hapten with EDC. The developed hapten coated icELISA can be used as a convenient quantitative tool for the sensitive and accurate monitoring DBP in water, plastic and cosmetic samples.

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Section: Discussionsupporting

confidence: 70%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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An Immunoassay for Dibutyl Phthalate Based on Direct Hapten Linkage to the Polystyrene Surface of Microtiter Plates

et al. 2011

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“…The immunization was done according to the protocol reported previously. 38 Each immunogen of BSA-P1 and BSA-P5 was injected into two female New Zealand white rabbits. The animals were bled around 3 months following the first immunization.…”

Section: Immunization Protocolmentioning

confidence: 99%

A sensitive and selective enzyme-linked immunosorbent assay for the analysis of para red in foods

Wang

Wei

et al. 2012

Analyst

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Para red is a synthetic dye and a potential genotoxic carcinogen. A hapten mimicking Para red structure was synthesized by introducing a carboxyl to the naphthol part of Para red and coupled to carrier protein to form an immunogen for the production of specific antibodies. A sensitive and selective enzyme-linked immunosorbent assay (ELISA) was developed for the detection of Para red in food samples. The limit of detection and inhibition half-maximum concentrations of Para red in phosphate buffered saline with 10% methanol were 0.06 and 2.2 ng mL(-1), respectively. Cross-reactivity values of the ELISA with the tested compounds including Sudan red I, II, III, IV, and G, sunset yellow, 2-naphthol, and 4-nitroaniline were ≤0.2%. This assay was used to determine Para red in tomato sauce, chilli sauce, chilli powder and sausage samples after ultrasonic extraction, cleanup and concentration steps. The average recoveries, repeatability (intraday extractions and analysis), and intra-laboratory reproducibility (interday extractions and analysis) were in the range 90-108%, 4-12% and 8-17%, respectively. This assay was compared to a high-performance liquid chromatographic method for 28 samples, displaying a good correlation (R(2) = 0.95). Para red residues in 53 real world samples determined by ELISA were below the limit of detection.

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“…For all the tested compounds, the cross reactivity values achieved were very low (52.5%) except for 2,3,4,6-tetrachlorophenol (5.6%) and 2,3,5,6-tetrachlorophenol (9.2%). The sensitivity and cross-reactivity of antibodies are related to the structure of immunizing haptens, coating antigen used, and physicochemical properties of ligand, including the minimum energy, threedimensional shape, electrostatic interaction, hydrophobic interaction, hydrogen bond (Xu et al 2007). The computational modeling may provide information useful in understanding cross reactivity.…”

Section: Antisera Specificitymentioning

confidence: 99%

Development of an enzyme-linked immunosorbent assay for the detection of pentachlorophenol residues in water samples

Xu¹,

Jing²,

Li³

et al. 2007

Food and Agricultural Immunology

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An indirect competitive enzyme-linked immunosorbent assay (ELISA) for pentachlorophenol (PCP) was developed with a polyclonal antiserum produced against a hapten, 4-(2,3,5,6-tetrachloro-4-hydroxyphenoxy)-butyric acid, conjugated to bovine serum albumin (BSA). The ELISA of PCP showed an IC 50 of 18 ng/ml and a limit of detection (LOD) of 0.9 ng/ml at the optimal concentration of NaCl (0.2 M) in the assay buffer. This assay was slightly affected by up to 10% (v/v) methanol or acetone. The IC 50 and A max values showed little change in a pH range of 5 and 7. Very low cross reactivities (52.5%) were observed for some structurally-related compounds, except for 2,3,4,6-tetrachlorophenol (5.6%) and 2,3,5,6-tetrachlorophenol (9.2%). The average recoveries of PCP from fortified water were in the range of 71Á118%. The results by GC and ELISA for the same fortified water samples showed non-significant difference (p 0.05). Finally, this ELISA was applied to monitor PCP in 100 river water samples and no PCP was detected. This ELISA is a good alternative tool for monitoring PCP residues in water samples.

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Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Pentachloronitrobenzene Residues in Environmental Samples

Cited by 10 publications

References 20 publications

An Immunoassay for Dibutyl Phthalate Based on Direct Hapten Linkage to the Polystyrene Surface of Microtiter Plates

An Immunoassay for Dibutyl Phthalate Based on Direct Hapten Linkage to the Polystyrene Surface of Microtiter Plates

A sensitive and selective enzyme-linked immunosorbent assay for the analysis of para red in foods

Development of an enzyme-linked immunosorbent assay for the detection of pentachlorophenol residues in water samples

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