title: Characterization of a 60-kDa Thermally Stable Antigenic Protein as a Marker for the Immunodetection of Bovine Plasma-Derived Food Ingredients.
abstract: A sandwich enzyme-linked immunosorbent assay (sELISA) based on 2 monoclonal antibodies (Bb3D6 and Bb6G12) that recognize a 60-kDa antigenic protein in bovine blood was previously developed for detecting bovine blood in animal feed for the prevention of mad cow disease. This study sought to establish the identity of this 60-kDa antigenic protein and consequently determine the suitability of the sELISA for detecting bovine plasma-derived food ingredients (BPFIs), which are widely used in dietary products without explicit labeling. Results from western blot confirmed the 60-kDa protein to be present in the plasma fraction of bovine blood. Further proteomic analyses involving 2-dimensional gel electrophoresis (2-D GE) and amino acid sequencing revealed the 60-kDa protein to be bovine serum albumin (BSA). The sELISA proved capable of detecting BPFIs in all the commercial dietary supplements tested, including those that were formulated with hydrolyzed BPFIs. The assay could also detect 0.01% and 0.5% of different BPFIs in spiked raw and cooked ground beef, respectively. This assay based on the detection of BSA therefore has the potential to become a valuable analytical tool to protect consumers who avoid consuming BPFIs for religious, health, or ethical reasons.
authors.affiliation: Dept. of Nutrition, Food and Exercise Sciences, Florida State Univ, Tallahassee, FL, 32306-1493, U.S.A.
authors.forename: Jack A
authors.initials: JA
authors.lastname: Ofori
authors.affiliation: Dept. of Nutrition, Food and Exercise Sciences, Florida State Univ, Tallahassee, FL, 32306-1493, U.S.A.
authors.forename: Yun-Hwa P
authors.initials: YH
authors.lastname: Hsieh
mesh_terms: D000821:Animal Feed; D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens; D015153:Blotting, Western; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005515:Food Labeling; D006358:Hot Temperature; D008460:Meat / Q000032:analysis*; D008970:Molecular Weight; D040901:Proteomics; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000737:chemistry / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D012710:Serum Albumin, Bovine
keywords: albumin; bovine; food ingredients; immunodetection; plasma