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

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.