Competitive enzyme-linked immunosorbent assay for quantitative detection of bovine blood in heat-processed meat and feed.
Animal blood is widely used as a functional additive in food and as a protein supplement in animal feed. Effective analytical methods are therefore needed to enforce labeling regulations for product quality control, as well as to address safety concerns. This study developed a monoclonal antibody (MAb)-based competitive enzyme-linked immunosorbent assay (ELISA) for quantitative detection of ruminant (bovine and ovine) blood in heat-processed meat and feedstuff. MAb BblH9 immunoglobulin G1, which recognizes a 12-kDa, thermostable, ruminant blood protein, was used as the detecting agent in the competitive ELISA. The immunoreactivity of MAb Bb1H9 was confirmed in both an indirect noncompetitive ELISA and a competitive ELISA, in which antigens are presented in immobilized form and free form, respectively. The competitive ELISA was optimized, and three spiked samples adulterated at 0 to 10% were tested to determine the detection limits of the optimized assay. Results showed that MAb Bb1H9 is specific to ruminant blood protein, with no cross-reaction with nonblood samples tested. The optimized competitive ELISA quantitatively detected heat-processed bovine blood over the tested range. The detection limit of the assay for bovine blood in beef (P < 0.001), bovine blood in porcine blood (P < 0.01), and bovine blood meal in soybean meal (P < 0.01) was found to be 0.5% in all cases. This MAb-based competitive ELISA is thus suitable for the sensitive and quantitative detection of ruminant blood proteins in heat processed meat and feed products.
Department of Nutrition, Food and Exercise Sciences, Florida State University, Tallahassee, Florida 32306-1493, USA.
Qinchun
Q
Rao
Yun-Hwa Peggy
YH
Hsieh
D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D000911:Antibodies, Monoclonal; D001798:Blood Proteins / Q000032:analysis*; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D001798:Blood Proteins