Immunochromatographic Detection of Myoglobin as a Specific Biomarker of Porcine Muscle Tissues in Meat Products

Zvereva, Elena A.; Byzova, Nadezhda A.; Hendrickson, Olga D.; Popravko, Demid S.; Belichenko, Ksenia A.; Dzantiev, Boris B.; Zherdev, Anatoly V.

doi:10.3390/app10217437

Cited by 20 publications

(9 citation statements)

References 32 publications

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“…As well as the commonly used 50 mM phosphate buffer, pH 7.4, with 0.1 M NaCl [ 46 ] was found to be unacceptable to exclude aggregation, it was replaced with 10 mM Tris-HCl, pH 8.8. The used stabilizing agent was BSA, because its effectiveness has been shown in a number of works [ 47 , 48 ]. In this media, the mAb−GNPs conjugate was stored for 10 months without changes in the properties of its colloid solution.…”

Section: Resultsmentioning

confidence: 99%

Development of Lateral Flow Test-System for the Immunoassay of Dibutyl Phthalate in Natural Waters

et al. 2022

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The use of a large amount of toxic synthetic materials leads to an increase in the pollution of environmental objects. Phthalates are compounds structurally related to esters of phthalic acid that are widely used in the manufacturing of synthetic packaging materials as plasticizers. Their danger is conditioned by leaching into the environment and penetrating into living organisms with negative consequences and effects on various organs and tissues. This work presents the first development of lateral flow immunoassay to detect dibutyl phthalate, one of the most common representatives of the phthalates group. To form a test zone, a hapten–protein conjugate was synthesized, and gold nanoparticles conjugated with antibodies to dibutyl phthalate were used as a detecting conjugate. The work includes the preparation of immunoreagents, selectivity investigation, and the study of the characteristics of the medium providing a reliable optical signal. Under the selected conditions for the analysis, the detection limit was 33.4 ng/mL, and the working range of the determined concentrations was from 42.4 to 1500 ng/mL. Time of the assay–15 min. The developed technique was successfully applied to detect dibutyl phthalate in natural waters with recovery rates from 75 to 115%.

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Section: Resultsmentioning

confidence: 99%

Development of Lateral Flow Test-System for the Immunoassay of Dibutyl Phthalate in Natural Waters

et al. 2022

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“…Accurate determination of Mbs in meat by ELISA requires full extraction of Mbs from meat. In previous reports of immunoassays for detecting pork, saline or buffer solutions have been used for the extraction of target proteins, − but the extraction efficiencies were low, especially in heated samples. As shown in Figure A, Mb of the heated sample was observed in the precipitate.…”

Section: Resultsmentioning

confidence: 99%

“…In contrast, immunoassays are easy and applicable for rapid detection of many samples with a simple sample preparation. Several immunoassays have been reported for the pork determination, in which α 1 -acid glycoprotein, troponin I, IgG, and myoglobin (Mb) are used as the target proteins. − However, these assays require appropriate standard substances for each food due to the differences in processing and accompanying protein denaturation and extraction efficiency. It would be impractical to attempt this for all foods.…”

Section: Introductionmentioning

confidence: 99%

Enzyme-Linked Immunosorbent Assay for Pork Determination in Raw and Heated Meats: Combination of Monoclonal Antibodies to Denatured Porcine Myoglobin and Sodium Dodecyl Sulfate Extraction

Yamasaki

Hirakawa

Momma

et al. 2021

ACS Food Sci. Technol.

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Several immunoassays for monitoring pork contamination have been developed, but they have limitations in accurate detection because they have been affected by protein denaturation and loss of extraction efficiency due to cooking. In this study, a sandwich enzyme-linked immunosorbent assay (s-ELISA) combined with an extraction method using sodium dodecyl sulfate (SDS) was developed for pork determination in raw and heated meats. To establish monoclonal antibodies (mAbs), SDS-denatured porcine myoglobin (Mb) and synthetic peptides with amino acid sequences of porcine Mb were used for immunization. The s-ELISA quantitatively detected the porcine Mb without cross-reaction to beef and chicken Mbs and lamb and goat meats. The 50% maximal effective concentration of porcine Mb for s-ELISA was 90 ng/mL, and the recoveries of porcine Mb spiked into raw and heated beef and chicken were 94−158%. This s-ELISA detected 1% (w/w) pork mixed with raw and heated beef. For the determination of Mbs in various parts of raw pork, including fatty bellies, the results correlated well with those obtained by high-performance liquid chromatography. Our s-ELISA could be available for authentication of meat products and prevention of mislabeling in pork products.

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“…As an effective molecular biomarker for meat authentication, myoglobin (MG) can be considered, being a muscle tissue protein characterized by different amino acid sequences for different animal species [27,28]. LFIAs of MG are described as molecular markers in some studies [29][30][31], where the authors successfully implemented species recognition of the meat sources.…”

Section: Introductionmentioning

confidence: 99%

Highly Sensitive Immunochromatographic Detection of Porcine Myoglobin as Biomarker for Meat Authentication Using Prussian Blue Nanozyme

Hendrickson,

Zvereva,

Dzantiev

et al. 2023

Foods

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This study was aimed at the sensitive immunodetection of porcine myoglobin (MG) as a species-specific biomarker in meat products. The enhanced lateral flow immunoassay (LFIA) was created in the sandwich format using monoclonal antibodies (Mab) with specificity to porcine MG and labeled by Prussian blue nanoparticles (PBNPs) as peroxidase-mimicking nanozymes. Signal amplification was provided by the colored product of oxidation catalyzed by the PBNPs. Several Mab–PBNP conjugates with different antibody loads were synthesized; the one that provided the best analytical characteristics of the LFIA was selected. Advanced optimization of the test system was carried out. As a result, the visual limit of detection (LOD) of MG was 1.5 ng/mL. Involvement of the catalytic nanozyme properties allowed the LOD to be decreased by ~9 times in comparison to the LFIA based on gold nanomarkers, and by ~27 times compared to the LFIA based on PBNP coloration. The assay time was 30 min, including catalytic enhancement. A simple technique of meat sample pre-treatment aimed at effective MG extraction and matrix disposal was proposed. The specificity of the LFIA towards the pork meat was demonstrated. The applicability of the created test system was shown by testing extracts obtained from finished meat products.

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Immunochromatographic Detection of Myoglobin as a Specific Biomarker of Porcine Muscle Tissues in Meat Products

Cited by 20 publications

References 32 publications

Development of Lateral Flow Test-System for the Immunoassay of Dibutyl Phthalate in Natural Waters

Development of Lateral Flow Test-System for the Immunoassay of Dibutyl Phthalate in Natural Waters

Enzyme-Linked Immunosorbent Assay for Pork Determination in Raw and Heated Meats: Combination of Monoclonal Antibodies to Denatured Porcine Myoglobin and Sodium Dodecyl Sulfate Extraction

Highly Sensitive Immunochromatographic Detection of Porcine Myoglobin as Biomarker for Meat Authentication Using Prussian Blue Nanozyme

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