Immunological detection of osteocalcin in meat and bone meal: a novel heat stable marker for the investigation of illegal feed adulteration

• Published in: Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment Chemistry, analysis, control, exposure & risk assessment, 2012-05, Vol.29 (5), p.716-726
• Main Authors: Kreuz, G., Zagon, J., Broll, H., Bernhardt, C., Linke, B., Lampen, A.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; animal feed; Animal Feed - analysis; Animals; Antibody Specificity; Biological and medical sciences; Biological Products - chemistry; Biomarkers; Bone and Bones - chemistry; bovine spongiform encephalopathy (BSE); Cattle; Diet; ELISA; Enzyme-Linked Immunosorbent Assay - veterinary; Epitopes; Europe; Feed and pet food industries; Food additives; Food Contamination; Food industries; Food Inspection - methods; Fundamental and applied biological sciences. Psychology; General aspects; Hot Temperature; Immunology; Industrial Waste - analysis; Limit of Detection; meat and bone meal (MBM); Meat Products - analysis; Meat-Packing Industry; Methods of analysis, processing and quality control, regulation, standards; Minerals - chemistry; Molecular Sequence Data; osteocalcin; Osteocalcin - analysis; Osteocalcin - chemistry; Peptides; processed animal proteins (PAPs); Protein Stability; Rodents; Sequence Alignment; species identification
• ISSN: 1944-0049; 1944-0057
• DOI: 10.1080/19440049.2011.645219

A sandwich ELISA was developed for the detection of bovine meat and bone meal (BMBM) in feed, based on polyclonal rabbit antibodies raised against the synthetic N-terminal amino acid sequence 1-9 (YLDHWLGAP) of bovine osteocalcin. To set up a sandwich ELISA pair, a commercial mouse monoclonal capture antibody binding to a highly conserved epitope in the mid-fragment of the peptide was employed. It is shown that the bone marker osteocalcin is immunologically well detectable in BMBM extracts obtained by a simple EDTA-based procedure even in a sample heated up to 145°C. Furthermore, a genus-specific restriction of the major specificity to cattle and horse was possible. The observed bi-specificity is consistent with theoretical predictions. The assay sensitivity with bovine osteocalcin of 1 ng was sufficient to enable the detection of 0.1% BMBM in compound plant feed or fish meal, for which no cross reaction was observed. In general the quantification of osteocalcin in extracts is possible using a standard curve procedure with pure bovine osteocalcin.