Production and Characterization of Monoclonal Antibodies to Pork Fat Protein

Introduction: Meat and food products adulterated with pork fat are a concern for Islamic consumers whose dietary laws prohibit consumption of pork meat. In addition, undeclared pork adulterated in meat products causes unexpected problems such as allergy and contamination of pathogenic bacteria and parasites. However no immunoassay for the rapid and simple detection of pork fat adulterated in meat and food products has been reported.

Purpose: The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to pork fat using thermal-stable soluble proteins as an immunogen and to detect pork fat by indirect ELISA.

Methods: Thermal-stable soluble protein extracted from pork fat was used as an immunogen. The mice showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained were confirmed by indirect ELISA and Western blot. Laboratory adulterated pork fat in other animal species (beef, chicken, duck, goat, turkey) were prepared and analyzed by indirect ELISA.

Results: Seven MAbs (2B8-3, 2B8-9, 2B8-20, 2B8-28, 2B8-31, 2B8-32 and 2B8-33) were developed. All MAbs were specific to pork fat without cross-reaction to pork meat and other animal species in the indirect ELISA and Western blot analyses. The ELISA assay can sensitively detect 1% pork fat protein in other animal species.

Significance: These results support that the application of ELISA could be used as rapid means to detect low levels of pork fat in meat and food products.