Bispecific Monoclonal Antibody-Based Multianalyte ELISA for Furaltadone Metabolite, Malachite Green, and Leucomalachite Green in Aquatic Products

Wang, Zewei; Wang, Hong; Shen, Yu-Dong; Li, Yongjun; Jiang, Dong; Xu, Zhen-Lin; Yang, Jin-Yi; Sun, Yuan; Xiao, Zhengguo

doi:10.1021/acs.jafc.6b03233

Cited by 44 publications

(11 citation statements)

References 40 publications

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“…The chemical mutagen scheme was performed as follows [28,45,46]: AFB 1 hybridoma cell line E 4 was cultured in HAT medium for 2 d before mutagenic treatment in a humidified 5% CO 2 incubator at 37 • C. The cells were washed twice with PBS and cultured in HT medium for 2 d to restore the cells to normal morphology and were cultured in HAT medium for 24 h. After washing the cells with PBS twice, the cells were mutagenized in different concentrations of MNNG (5.0 µg/mL, 10.0 µg/mL, 20.0 µg/mL, and 40.0 µg/mL). The mutagenic treatment time was 2 h, 4 h, 6 h and 10 h. At the same time, the control group was cultured in 1‰ DMSO complete medium (RPMI-1640 culture media containing 20% fetal bovine serum) without MNNG.…”

Section: Mutagenesis Of Hgprt and Tk Deficient Hybridoma Cell Linesmentioning

confidence: 99%

Preparation of an Immunoaffinity Column Based on Bispecific Monoclonal Antibody for Aflatoxin B1 and Ochratoxin A Detection Combined with ic-ELISA

Wang

et al. 2022

Foods

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A novel and efficient immunoaffinity column (IAC) based on bispecific monoclonal antibody (BsMAb) recognizing aflatoxin B1 (AFB1) and ochratoxin A (OTA) was prepared and applied in simultaneous extraction of AFB1 and OTA from food samples and detection of AFB1/OTA combined with ic-ELISA (indirect competitive ELISA). Two deficient cell lines, hypoxanthine guanine phosphoribosyl-transferase (HGPRT) deficient anti-AFB1 hybridoma cell line and thymidine kinase (TK) deficient anti-OTA hybridoma cell line, were fused to generate a hybrid-hybridoma producing BsMAb against AFB1 and OTA. The subtype of the BsMAb was IgG1 via mouse antibody isotyping kit test. The purity and molecular weight of BsMAb were confirmed by SDS-PAGE method. The cross-reaction rate with AFB2 was 37%, with AFG1 15%, with AFM1 48%, with AFM2 10%, and with OTB 36%. Negligible cross-reaction was observed with other tested compounds. The affinity constant (Ka) was determined by ELISA. The Ka (AFB1) and Ka (OTA) was 2.43 × 108 L/mol and 1.57 × 108 L/mol, respectively. Then the anti-AFB1/OTA BsMAb was coupled with CNBr-Sepharose, and an AFB1/OTA IAC was prepared. The coupling time and elution conditions of IAC were optimized. The coupling time was 1 h with 90% coupling rate, the eluent was methanol–water (60:40, v:v, pH 2.3) containing 1 mol/L NaCl, and the eluent volume was 4 mL. The column capacities of AFB1 and OTA were 165.0 ng and 171.3 ng, respectively. After seven times of repeated use, the preservation rates of column capacity for AFB1 and OTA were 69.3% and 68.0%, respectively. The ic-ELISA for AFB1 and OTA were applied combined with IAC. The IC50 (50% inhibiting concentration) of AFB1 was 0.027 ng/mL, the limit of detection (LOD) was 0.004 ng/mL (0.032 µg/kg), and the linear range was 0.006 ng/mL~0.119 ng/mL. The IC50 of OTA was 0.878 ng/mL, the LOD was 0.126 ng/mL (1.008 µg/kg), and the linear range was 0.259 ng/mL~6.178 ng/mL. Under optimum conditions, corn and wheat samples were pretreated with AFB1-OTA IAC. The recovery rates of AFB1 and OTA were 95.4%~105.0% with ic-ELISA, and the correlations between the detection results and LC-MS were above 0.9. The developed IAC combined with ic-ELISA is reliable and could be applied to the detection of AFB1 and OTA in grains.

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Section: Mutagenesis Of Hgprt and Tk Deficient Hybridoma Cell Linesmentioning

confidence: 99%

Preparation of an Immunoaffinity Column Based on Bispecific Monoclonal Antibody for Aflatoxin B1 and Ochratoxin A Detection Combined with ic-ELISA

Wang

et al. 2022

Foods

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“…Cell fusion was carried out in reference to a previously-described process (Wang et al, 2015(Wang et al, , 2016. The spleen cells and the mouse myeloma cells SP2/0 were fused using PEG 1450.…”

Section: Preparation Of the Monoclonal Antibodymentioning

confidence: 99%

Ultrasensitive and simultaneous detection of 6 nonsteroidal anti-inflammatory drugs by colloidal gold strip sensor

Lü

Kuang

et al. 2021

Journal of Dairy Science

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In this work, an oxicam group-selective monoclonal antibody against 6 nonsteroidal anti-inflammatory drugs (NSAID; meloxicam, lornoxicam, piroxicam, sudoxicam, droxicam, and tenoxicam) was prepared. Also, a spacer arm with carboxyl group was derived at the hydroxyl of meloxicam to generate the meloxicam hapten. The half-maximal inhibitory concentrations (IC 50 ) were, respectively, 0.31 ng/mL for meloxicam, 0.49 ng/mL for lornoxicam, 2.90 ng/mL for piroxicam, 1.95 ng/mL for sudoxicam, 3.08 ng/mL for droxicam, and 5.36 ng/mL for tenoxicam. A colloidal gold immunochromatographic strip based on the monoclonal antibody was developed for the detection of these 6 NSAID in milk. The results could be obtained by the naked eye in 10 min, and the cut-off values and the visual limits of detection in real samples were 5, 5, 10, 10, 25, and 25 ng/mL, and 0.25, 1, 0.5, 0.5, 1, and 1 ng/ mL, respectively. This immunochromatopgraphic strip is a suitable tool for on-site detection and screening of oxicam NSAID in milk samples.

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“…Several multi-analyte immunoassays for the screening of pesticide and veterinary drug residues have been developed using bispecific antibodies. A bispecific antibody-based icELISA method was developed for the detection of 5-morpholinomethyl-3-amino-2-oxazolidone and leucomalachite green in aquatic products [52]. The LODs for 5-morpholinomethyl-3-amino-2-oxazolidone and leucomalachite green were 0.2 and 4.8 ng/mL, respectively.…”

Section: Preparation and Application Of Bispecific Antibodiesmentioning

confidence: 99%

Rapid Multi-Residue Detection Methods for Pesticides and Veterinary Drugs

et al. 2020

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The excessive use or abuse of pesticides and veterinary drugs leads to residues in food, which can threaten human health. Therefore, there is an extremely urgent need for multi-analyte analysis techniques for the detection of pesticide and veterinary drug residues, which can be applied as screening techniques for food safety monitoring and detection. Recent developments related to rapid multi-residue detection methods for pesticide and veterinary drug residues are reviewed herein. Methods based on different recognition elements or the inherent characteristics of pesticides and veterinary drugs are described in detail. The preparation and application of three broadly specific recognition elements—antibodies, aptamers, and molecular imprinted polymers—are summarized. Furthermore, enzymatic inhibition-based sensors, near-infrared spectroscopy, and SERS spectroscopy based on the inherent characteristics are also discussed. The aim of this review is to provide a useful reference for the further development of rapid multi-analyte analysis of pesticide and veterinary drug residues.

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Bispecific Monoclonal Antibody-Based Multianalyte ELISA for Furaltadone Metabolite, Malachite Green, and Leucomalachite Green in Aquatic Products

Cited by 44 publications

References 40 publications

Preparation of an Immunoaffinity Column Based on Bispecific Monoclonal Antibody for Aflatoxin B1 and Ochratoxin A Detection Combined with ic-ELISA

Preparation of an Immunoaffinity Column Based on Bispecific Monoclonal Antibody for Aflatoxin B1 and Ochratoxin A Detection Combined with ic-ELISA

Ultrasensitive and simultaneous detection of 6 nonsteroidal anti-inflammatory drugs by colloidal gold strip sensor

Rapid Multi-Residue Detection Methods for Pesticides and Veterinary Drugs

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