Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies

A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies directed against a synthetic peptide with an amino-acid sequence related to the C-terminus of bovine myoglobin and the whole molecule of sodium dodecyl sulphate (SDS)-denatured bovine myoglobin was...

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Autores principales: YAMAMOTO, Takayuki, KATO, Masatoshi, ENDO, Kiwamu, KOTOURA, Satoshi, TAKEDA, Zenya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710716/
https://www.ncbi.nlm.nih.gov/pubmed/26166832
http://dx.doi.org/10.1292/jvms.15-0078
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author YAMAMOTO, Takayuki
KATO, Masatoshi
ENDO, Kiwamu
KOTOURA, Satoshi
TAKEDA, Zenya
author_facet YAMAMOTO, Takayuki
KATO, Masatoshi
ENDO, Kiwamu
KOTOURA, Satoshi
TAKEDA, Zenya
author_sort YAMAMOTO, Takayuki
collection PubMed
description A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies directed against a synthetic peptide with an amino-acid sequence related to the C-terminus of bovine myoglobin and the whole molecule of sodium dodecyl sulphate (SDS)-denatured bovine myoglobin was adapted for detecting bovine myoglobin in contaminated feeds. The ELISA employed bovine meat extract of a known myoglobin concentration as a calibration standard and had an limit of detection (LOD) of 3.54 ng/ml and an limit of quantification (LOQ) of 11.0 ng/ml corresponding to 0.022% and 0.067% (wt/wt) bovine meat-and-bone-meal (MBM) mixed in 20-fold-diluted feed extracts, respectively. A cut-off threshold of 20.6 ng/ml bovine myoglobin was set to simplify ELISA and facilitate quick assessment of test results without a tedious calibration process. The ELISA was able to detect bovine MBM in artificially prepared model feeds, mixed botanical feeds, mixed botanical feeds with skimmed milk, fish meal, pork meal and pork/chicken meal at 0.1% (wt/wt). It was also able to detect sheep MBM in test feeds, but showed no reactivity to swine MBM, chicken MBM, skimmed milk or gelatine of bovine origin. The advantages of this method are the quick and easy extraction protocol of proteins from test feeds, using 100 mM sodium sulphide and 0.6% sodium dodecyl sulphate in the extraction solution and the effective detection of bovine and sheep MBM at 0.1% (wt/wt).
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spelling pubmed-47107162016-01-14 Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies YAMAMOTO, Takayuki KATO, Masatoshi ENDO, Kiwamu KOTOURA, Satoshi TAKEDA, Zenya J Vet Med Sci Biochemistry A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies directed against a synthetic peptide with an amino-acid sequence related to the C-terminus of bovine myoglobin and the whole molecule of sodium dodecyl sulphate (SDS)-denatured bovine myoglobin was adapted for detecting bovine myoglobin in contaminated feeds. The ELISA employed bovine meat extract of a known myoglobin concentration as a calibration standard and had an limit of detection (LOD) of 3.54 ng/ml and an limit of quantification (LOQ) of 11.0 ng/ml corresponding to 0.022% and 0.067% (wt/wt) bovine meat-and-bone-meal (MBM) mixed in 20-fold-diluted feed extracts, respectively. A cut-off threshold of 20.6 ng/ml bovine myoglobin was set to simplify ELISA and facilitate quick assessment of test results without a tedious calibration process. The ELISA was able to detect bovine MBM in artificially prepared model feeds, mixed botanical feeds, mixed botanical feeds with skimmed milk, fish meal, pork meal and pork/chicken meal at 0.1% (wt/wt). It was also able to detect sheep MBM in test feeds, but showed no reactivity to swine MBM, chicken MBM, skimmed milk or gelatine of bovine origin. The advantages of this method are the quick and easy extraction protocol of proteins from test feeds, using 100 mM sodium sulphide and 0.6% sodium dodecyl sulphate in the extraction solution and the effective detection of bovine and sheep MBM at 0.1% (wt/wt). The Japanese Society of Veterinary Science 2015-07-12 2015-12 /pmc/articles/PMC4710716/ /pubmed/26166832 http://dx.doi.org/10.1292/jvms.15-0078 Text en ©2015 The Japanese Society of Veterinary Science http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License.
spellingShingle Biochemistry
YAMAMOTO, Takayuki
KATO, Masatoshi
ENDO, Kiwamu
KOTOURA, Satoshi
TAKEDA, Zenya
Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies
title Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies
title_full Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies
title_fullStr Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies
title_full_unstemmed Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies
title_short Detection of ruminant meat and bone meal in feeds by sandwich ELISA with monoclonal antibodies
title_sort detection of ruminant meat and bone meal in feeds by sandwich elisa with monoclonal antibodies
topic Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4710716/
https://www.ncbi.nlm.nih.gov/pubmed/26166832
http://dx.doi.org/10.1292/jvms.15-0078
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