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Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products
Probiotic bacteria have a wide range of applications in veterinary and human therapeutics. Inactivated probiotics are complex samples and quality control (QC) should measure as many molecular features as possible. Capillary electrophoresis coupled to mass spectrometry (CE/MS) has been used as a mult...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3686750/ https://www.ncbi.nlm.nih.gov/pubmed/23840518 http://dx.doi.org/10.1371/journal.pone.0066682 |
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author | Klein, Günter Schanstra, Joost P. Hoffmann, Janosch Mischak, Harald Siwy, Justyna Zimmermann, Kurt |
author_facet | Klein, Günter Schanstra, Joost P. Hoffmann, Janosch Mischak, Harald Siwy, Justyna Zimmermann, Kurt |
author_sort | Klein, Günter |
collection | PubMed |
description | Probiotic bacteria have a wide range of applications in veterinary and human therapeutics. Inactivated probiotics are complex samples and quality control (QC) should measure as many molecular features as possible. Capillary electrophoresis coupled to mass spectrometry (CE/MS) has been used as a multidimensional and high throughput method for the identification and validation of biomarkers of disease in complex biological samples such as biofluids. In this study we evaluate the suitability of CE/MS to measure the consistency of different lots of the probiotic formulation Pro-Symbioflor which is a bacterial lysate of heat-inactivated Escherichia coli and Enterococcus faecalis. Over 5000 peptides were detected by CE/MS in 5 different lots of the bacterial lysate and in a sample of culture medium. 71 to 75% of the total peptide content was identical in all lots. This percentage increased to 87–89% when allowing the absence of a peptide in one of the 5 samples. These results, based on over 2000 peptides, suggest high similarity of the 5 different lots. Sequence analysis identified peptides of both E. coli and E. faecalis and peptides originating from the culture medium, thus confirming the presence of the strains in the formulation. Ontology analysis suggested that the majority of the peptides identified for E. coli originated from the cell membrane or the fimbrium, while peptides identified for E. faecalis were enriched for peptides originating from the cytoplasm. The bacterial lysate peptides as a whole are recognised as highly conserved molecular patterns by the innate immune system as microbe associated molecular pattern (MAMP). Sequence analysis also identified the presence of soybean, yeast and casein protein fragments that are part of the formulation of the culture medium. In conclusion CE/MS seems an appropriate QC tool to analyze complex biological products such as inactivated probiotic formulations and allows determining the similarity between lots. |
format | Online Article Text |
id | pubmed-3686750 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-36867502013-07-09 Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products Klein, Günter Schanstra, Joost P. Hoffmann, Janosch Mischak, Harald Siwy, Justyna Zimmermann, Kurt PLoS One Research Article Probiotic bacteria have a wide range of applications in veterinary and human therapeutics. Inactivated probiotics are complex samples and quality control (QC) should measure as many molecular features as possible. Capillary electrophoresis coupled to mass spectrometry (CE/MS) has been used as a multidimensional and high throughput method for the identification and validation of biomarkers of disease in complex biological samples such as biofluids. In this study we evaluate the suitability of CE/MS to measure the consistency of different lots of the probiotic formulation Pro-Symbioflor which is a bacterial lysate of heat-inactivated Escherichia coli and Enterococcus faecalis. Over 5000 peptides were detected by CE/MS in 5 different lots of the bacterial lysate and in a sample of culture medium. 71 to 75% of the total peptide content was identical in all lots. This percentage increased to 87–89% when allowing the absence of a peptide in one of the 5 samples. These results, based on over 2000 peptides, suggest high similarity of the 5 different lots. Sequence analysis identified peptides of both E. coli and E. faecalis and peptides originating from the culture medium, thus confirming the presence of the strains in the formulation. Ontology analysis suggested that the majority of the peptides identified for E. coli originated from the cell membrane or the fimbrium, while peptides identified for E. faecalis were enriched for peptides originating from the cytoplasm. The bacterial lysate peptides as a whole are recognised as highly conserved molecular patterns by the innate immune system as microbe associated molecular pattern (MAMP). Sequence analysis also identified the presence of soybean, yeast and casein protein fragments that are part of the formulation of the culture medium. In conclusion CE/MS seems an appropriate QC tool to analyze complex biological products such as inactivated probiotic formulations and allows determining the similarity between lots. Public Library of Science 2013-06-19 /pmc/articles/PMC3686750/ /pubmed/23840518 http://dx.doi.org/10.1371/journal.pone.0066682 Text en © 2013 Klein et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Klein, Günter Schanstra, Joost P. Hoffmann, Janosch Mischak, Harald Siwy, Justyna Zimmermann, Kurt Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products |
title | Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products |
title_full | Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products |
title_fullStr | Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products |
title_full_unstemmed | Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products |
title_short | Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products |
title_sort | proteomics as a quality control tool of pharmaceutical probiotic bacterial lysate products |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3686750/ https://www.ncbi.nlm.nih.gov/pubmed/23840518 http://dx.doi.org/10.1371/journal.pone.0066682 |
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