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Proteomic Analysis and Label-Free Quantification of the Large Clostridium difficile Toxins
Joint Authors
Williamson, Yulanda M.
Solano, Maria I.
Woolfitt, Adrian R.
Blake, Thomas A.
Moura, Hercules
Wagner, Glauber
Barr, John R.
Terilli, Rebecca R.
Source
International Journal of Proteomics
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-08-27
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Natural & Life Sciences (Multidisciplinary)
Biology
Abstract EN
Clostridium difficile is the leading cause of antibiotic-associated diarrhea in hospitals worldwide, due to hypervirulent epidemic strains with the ability to produce increased quantities of the large toxins TcdA and TcdB.
Unfortunately, accurate quantification of TcdA and TcdB from different toxinotypes using small samples has not yet been reported.
In the present study, we quantify C.
difficile toxins in <0.1 mL of culture filtrate by quantitative label-free mass spectrometry (MS) using data-independent analysis (MSE).
In addition, analyses of both purified TcdA and TcdB as well as a standard culture filtrate were performed using gel-based and gel-independent proteomic platforms.
Gel-based proteomic analysis was then used to generate basic information on toxin integrity and provided sequence confirmation.
Gel-independent in-solution digestion of both toxins using five different proteolytic enzymes with MS analysis generated broad amino acid sequence coverage (91% for TcdA and 95% for TcdB).
Proteomic analysis of a culture filtrate identified a total of 101 proteins, among them TcdA, TcdB, and S-layer proteins.
American Psychological Association (APA)
Moura, Hercules& Terilli, Rebecca R.& Woolfitt, Adrian R.& Williamson, Yulanda M.& Wagner, Glauber& Blake, Thomas A.…[et al.]. 2013. Proteomic Analysis and Label-Free Quantification of the Large Clostridium difficile Toxins. International Journal of Proteomics،Vol. 2013, no. 2013, pp.1-10.
https://search.emarefa.net/detail/BIM-461070
Modern Language Association (MLA)
Moura, Hercules…[et al.]. Proteomic Analysis and Label-Free Quantification of the Large Clostridium difficile Toxins. International Journal of Proteomics No. 2013 (2013), pp.1-10.
https://search.emarefa.net/detail/BIM-461070
American Medical Association (AMA)
Moura, Hercules& Terilli, Rebecca R.& Woolfitt, Adrian R.& Williamson, Yulanda M.& Wagner, Glauber& Blake, Thomas A.…[et al.]. Proteomic Analysis and Label-Free Quantification of the Large Clostridium difficile Toxins. International Journal of Proteomics. 2013. Vol. 2013, no. 2013, pp.1-10.
https://search.emarefa.net/detail/BIM-461070
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-461070