Monoclonal antibody-based ELISA for the quantification of porcine hemoglobin in meat products

Jiang, Xingyi; Fuller, D.T.; Hsieh, Yun-Hwa Peggy; Rao, Qinchun

doi:10.1016/j.foodchem.2018.01.032

Cited by 33 publications

(14 citation statements)

References 24 publications

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“…Various methods including protein- and DNA-based assays have been developed to detect meat authenticity [ 10 , 11 , 12 , 13 , 14 ]. However, the identification of meat species mainly adopts DNA-based nucleic acid detection methods, of which the most widely used methods are the polymerase chain reaction (PCR) based methods [ 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Recombinase Polymerase Amplification Based Multiplex Lateral Flow Dipstick for Fast Identification of Duck Ingredient in Adulterated Beef

Zhang

Zhou

et al. 2020

Animals

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Meat adulteration has become a global social problem. In order to protect consumers from meat adulteration, several methods have been developed to identify meat species. However, the conventional methods are labor-intensive, time-consuming and require instruments. In the present study, a rapid and visual method based on recombinase polymerase amplification (RPA) and multiplex lateral flow dipstick (MLFD) was developed to detect duck ingredient in adulterated beef. Using recombinase and strand displacement polymerase enable RPA to amplify different double-labeled DNA amplicons at room temperature, which can be further detected by MLFD. The whole reaction process can be finished within 35 min, and the results can be determined by naked eyes. As low as 5% of duck ingredient in adulterated beef can be easily measured. Moreover, we confirmed that our new method held good potential in the detection of commercially processed meat samples. In conclusion, this study reported a useful animal derived meat adulteration detection method, which have potential application in future.

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Section: Introductionmentioning

confidence: 99%

Recombinase Polymerase Amplification Based Multiplex Lateral Flow Dipstick for Fast Identification of Duck Ingredient in Adulterated Beef

Zhang

Zhou

et al. 2020

Animals

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“…Blood, meat protein extracts, and positive controls (P Hb , mouse anti-P Hb mAb IgG, and rabbit anti-P Hb pAb IgG) were separated using a non-reducing sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE, 4% stacking gel and 15% separating gel) according to Jiang et al [27] with modifications. Briefly, electrophoretically separated protein bands on the gel were transferred onto a 0.45-µm nitrocellulose membrane (Thermo Scientific).…”

Section: Western Blotmentioning

confidence: 99%

Non-Specific Binding and Cross-Reaction of ELISA: A Case Study of Porcine Hemoglobin Detection

Jiang

Albo

et al. 2021

Foods

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Different types of enzyme-linked immunosorbent assays (ELISA) have been widely used to control food safety and quality. To develop an accurate and reproducible ELISA, false immunodetection results caused by non-specific binding (NSB) and cross-reaction must be prevented. During the case study of sandwich ELISA development for the detection of porcine hemoglobin (PHb), several critical factors leading to NSB and cross-reaction were found. First, to reduce the NSB of the target analyte, the selection of microplate and blocker was discussed. Second, cross-reactions between enzyme-labeled secondary antibodies and sample proteins were demonstrated. In addition, the function of (3-aminopropyl)triethoxysilane (APTES) was evaluated. Overall, this study highlights the essence of both antibody and assay validation to minimize any false-positive/negative immunodetection results.

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“…As for IC 50 , it was defined as the concentration that the combination of antibody to IM-OVA is inhibited by 50%. The procedure of ic-ELISA was developed based on the former description (Jiang et al, 2018). The optimal density of the IM-OVA was 0.31 µg/ml and mAb was 0.41 µg/ml.…”

Section: Characterization Of the Ic-elisamentioning

confidence: 99%

Immunochromatographic assay based on high‐affine monoclonal antibody for the detection of imidocarb in milk

Chen

Sun

et al. 2021

Journal of Food Science

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Imidocarb (IM) is an antiprotozoal agent, which is mainly used to treat babesiosis and piroplasmosis for animals. However, overdose or improbable utilization cause IM residues in animal origin products, which would be harmful to human health. Here, a monoclonal antibody (mAb) against IM with extremely sensitive and specific features has been successfully prepared from a novel immunogen synthesized by virtue of the active ester method. The concentration of half-maximal inhibition (IC 50 ) of the mAb was 0.074 ng/ml and the affinity constant was 4.58 × 10 10 L/mol. On the basis of this condition, an immunochromatographic strip (ICS) is proposed that could be applied in milk samples to test IM rapidly. For the ICS, the visual detection limit (cut-off value) was 5 ng/ml, IC 50 was 0.4 ng/ml, the limit of detection (LOD) was 0.078 ng/ml, the linear detection scope was 0.117 to 1.37 ng/ml. The recovery rates ranged from 88.83% to 91.47% and coefficients of variation (CV) were in the spectrum of 7.31% to 9.43%. In general, the recommended test strip provided an exceedingly simple and reliable detection method for quickly testing the IM. Practical Application: In our joint efforts, an extremely sensitive monoclonal antibody against imidocarb was obtained and a test strip for the rapid detection of imidocarb in milk was developed. The developed method could be applied to the field and provided great potential for analytical of imidocarb in other matrixes.

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Monoclonal antibody-based ELISA for the quantification of porcine hemoglobin in meat products

Cited by 33 publications

References 24 publications

Recombinase Polymerase Amplification Based Multiplex Lateral Flow Dipstick for Fast Identification of Duck Ingredient in Adulterated Beef

Recombinase Polymerase Amplification Based Multiplex Lateral Flow Dipstick for Fast Identification of Duck Ingredient in Adulterated Beef

Non-Specific Binding and Cross-Reaction of ELISA: A Case Study of Porcine Hemoglobin Detection

Immunochromatographic assay based on high‐affine monoclonal antibody for the detection of imidocarb in milk

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