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

Abstract

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.

Porous carbon‐based electrochemical platform for direct electron transfer in myoglobin

“…Some reports employed different morphological materials along with denaturants [9, 25–30] and redox mediators [31]. The denaturant agent such as urea and guanidine hydrochloride [32] were employed in order to open up the pocket to expose the heme center for direct electron transfer.…”

Rapid detection of duck ingredient in adulterated foods by isothermal recombinase polymerase amplification assays

“…However, their application in the food industry for routine analysis is rarely discussed. Currently, the primary techniques for meat identification and adulteration are protein-based methods ( Mandli et al, 2018 , Yamasaki et al, 2021 ) and DNA-based methods ( Chen et al, 2015 , Liu et al, 2019 ). Polymerase Chain Reaction (PCR)-based methods and enzyme-linked immunosorbent assay (ELISA) are the two main methods to detect meat adulteration ( Konduru et al, 2021 , Yin et al, 2020 ), and the former was extensively studied in the literature.…”

Zero-Biased Photoelectrochemical Detection of Cardiac Biomarker Myoglobin Based on CdSeS/ZnS Quantum Dots and Barium Titanate Perovskite

“…Thus, a specific, highly sensitive, and early detection of Mb in the blood is essential for diagnosing AMI [ 2 , 4 , 5 , 6 ]. In this sense, many methods are reported in the literature for the detection of Mb, such as immunoenzymatic assay [ 7 , 8 ], fluorescence [ 9 , 10 ], surface plasmon resonance [ 11 , 12 ], mass spectrometry [ 13 , 14 ], and liquid chromatography [ 15 , 16 ]. However, although such methods have a higher selectivity for Mb quantification, they are very expensive and require long sample preparation and analysis.…”