Identification of Microorganisms by Mass Spectrometry / Edition 1

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A multidisciplinary approach to understanding the fundamentals of mass spectrometry for bacterial analysis

From chemotaxonomy to characterization of targeted proteins, Identification of Microorganisms by Mass Spectrometry provides an overview of both well-established and cutting-edge mass spectrometry techniques for identifying microorganisms. A vital tool for microbiologists, health professionals, and analytical chemists, the text is designed to help scientists select the most effective techniques for use in biomedical, biochemical, pharmaceutical, and bioterror defense applications.

Since microbiological applications of mass spectrometry require a basic understanding of both microbiology and analytical chemistry, the editors have incorporated material from both disciplines so that readers from either field will come to understand the necessary principles of the other. Featuring contributions from some of the most recognized experts in both fields, this volume provides specific examples of fundamental methods as well as approaches developed in the last decade, including:
* Metastable atom bombardment pyrolysis mass spectrometry
* Matrix-assisted laser desorption/ionization mass spectrometry (MALDI)
* MALDI time-of-flight mass spectrometry (MALDI-TOF MS) of intact bacteria
* High-resolution Fourier transform mass spectrometry (FTMS)
* Electrospray ionization (ESI) mass spectrometry

Identification of Microorganisms by Mass Spectrometry represents the most comprehensive and up-to-date work on the topic currently available. It is liberally illustrated with figures and tables and covers every aspect of spectrometric identification of microorganisms, including experimental procedures, various means of sample preparation, data analysis, and interpretation of complex mass spectral data.

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Editorial Reviews

From the Publisher
"...provides up-to-date material on an important field that isbound to continue to grow, and is highly recommended." (TheQuarterly Review of Biology, September 2006)

"...very well-written and updated, and it should serve a needfor both newcomers and experienced researchers in the field."(Journal of the American Society for Mass Spectrometry, July2006)

"…this book fills a gap in teaching microbiologists andanalytical chemists alike how to use the fundamentals of eachothers’ fields." (Chimie Nouvelle, June 2007)

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Product Details

Meet the Author

CHARLES L. WILKINS, PhD, is a Distinguished Professor in theDepartment of Chemistry and Biochemistry at the University ofArkansas. He is the author or coauthor of over 250 scientificpublications and has received numerous awards for his research.

JACKSON O. LAY, Jr., PhD, is the Director of the ArkansasStatewide Mass Spectrometry Facility at the University of Arkansas,and the author or coauthor of more than 100 publications in thefield. In addition to his work with bacteria, he has developed massspectrometry methods for cancer research and toxicology.

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Table of Contents



1. Cultural, Serological and Genetic Methods for Identificationof Bacteria (John B. Sutherland and Fatemeh Rafii).

1.1 Introduction.

1.2 Identification of bacteria by cultural methods.

1.3 Identification of bacteria by serological methods.

1.4 Identification of bacteria by genetic methods.

1.5 Other methods used for bacterial characterization.

1.6 Conclusions.

1.7 Acknowledgments and Disclaimers.

1.8 References.

2. Mass Spectrometry: Identification and Biodetection, LessonsLearned and Future Developments (Alvin Fox).

2.1 Introduction.

2.1.1 Analysis of Fatty Acid and Sugar Monomers using GC-FID,GC-MS and GC-MS-MS.

2.1.2 2.2 Analysis of PCR products using PCR, PCR-MS andPCR-MS-MS.

2.1.3 Analysis of proteins using MALDI-TOF MS.

2.1.4 Chemical markers for protein-based identification orbiodetection.

2.1.5 Conclusions.

2.1.6 References.

3. An Introduction to Maldi TOF MS Analysis of WholeBacteria (Rohana Liyanage and Jackson O. Lay, Jr.).

3.1. Introduction.

3.2. Mass Spectrometry and Time-of-Flight MS.

3.3. Matrix Assisted Laser Desorption Ionization.

3.4. MALDI/TOF Mass Spectrometry.

3.5. MALDI TOF and Bacterial Identification.

3.6. Conclusions.

3.7. References.

4. The Development of the Block II Chemical Biological MassSpectrometer (Wayne H. Griest and Stephen A. Lammert.

4.1. Introduction.

4.2. Development History and Design Philosophy.

4.3. Requirements and Specifications.

4.4. Performance Testing.

4.5. Conclusions.

4.6. Acknowledgements.

4.7 References.

5. Method Reproducibility and Spectral Library Assembly forRapid Bacterial Characterization by Metastable Atom BombardmentPyrolysis Mass Spectrometry (Jon G. Wilkes, Gary Miertschin, ToddEschler, Les Hosey, Fatemeh Rafii, Larry Rushing, Dan A. Buzatu,and Michel J. Bertrand).

5.1. Introduction.

5.2. Sample Preparation for Rapid, Reproducible CellCulture.

5.3. Analytical instrumentation for sensitive detection andspectral reproducibility.

5.4. Spectral library assembly.

5.5. Pattern Recognition Methods for Objectively ClassifyingBacteria.

5.6. Conclusions.

5.7. Acknowledgement and Disclaimer.

5.8. References.

6. MALDI TOF Mass Spectrometry of Intact Bacteria (Jackson O.Lay, Jr., and Rohana Liyanage).

6.1. Introduction.

6.2. MALDI MS of Cellular Extracts.

6.3. Taxonomy: From Isolates to Whole Cell MALDI.

6.4 Whole Cell MALDI MS.

6.5 Biology Based Changes in Whole Cell MALDI Spectra.

6.6 Analysis of Mixtures.

6.7 Experimental Approaches.

6.8 Identification of Protein Markers.

6.9 Analysis of Target Proteins.

6.10 Analysis of Clinical Isolates.

6.11 Conclusions.

6.12 References.

7. Development of Spectral Pattern Matching Approaches toMatrix-Assisted Laser Desorption/Ionization Time-of-Flight MassSpectrometry for Bacterial Identification (Kristin H. Jarman andKaren L. Wahl).

7.1. Introduction.

7.2. MALDI-MS Signature Library Construction andIdentification.

7.3.Future Directions.

7.4. References.

8. Studies of Malaria by Mass Spectrometry (Plamen A.Demirev).

8.1. Introduction.

8.2. Plasmodium in Red Blood Cells.

8.3. Experimental Protocols for LDMS Detection of Malaria.

8.4. Malaria Detection by Laser Desorption MassSpectrometry.

8.5. MS-Based Proteomics of the Plasmodium Parasite.

8.6. Conclusions.

8.7. Acknowledgements.

8.8. References.

9. Bacterial Strain Differentiation by Mass Spectrometry (RandyJ. Arnold, Jonathan A. Karty and James P. Reilly).

9.1. Introduction.

9.2. Analysis of Cellular Proteins by Mass Spectrometry.

9.3. Application of MALDI-TOF to Bacteria Identification.

9.4. Conclusions.

9.5. Acknowledgments.

9.6. References.

10. Bacterial Protein Biomarker Discovery: A Focused Approach toDeveloping Molecular Based Identification Systems (Tracie L.Williams, Steven R. Monday, and Steven M. Musser).

10.1. Introduction.

10.2. Protein Extraction Methods.

10.3. Mass Spectrometry.

10.4. Automating the Process.

10.5. Collecting and Sequencing Proteins.

10.6. Conclusions.

11. High Throughput Microbial Characterisations UsingElectrospray Ionisation Mass Spectrometry and Its Role inFunctional Genomics (Seetharaman Vaidyanathan and RoystonGoodacre).

11.1. Introduction.

11.2. Microbial characterisations beyond the genomic level:Functional genomics.

11.3. Electrospray (ionization) mass spectrometry (ESMS).

11.4. ESMS of Microbes.

11.5. Direct infusion ESMS of crude cell extracts forhigh-throughput characterizations - metabolic fingerprinting andfootprinting.

11.6. Conclusions.

11.7. Acknowledgements.

11.8. References.

12. Bioinformatics for Flexibility, Reliability and MixtureAnalysis of Intact Microorganisms (Catherine Fenselau and PatrickPribil).

12.1. Introduction.

12.2. Library Matching.

12.3. Machine Learning.

12.4. Bioinformatics.

12.5. Protein Molecular Masses.

12.6. Protein Maps.

12.7 Microsequences from Peptides and Proteins.

12.8 Remaining Challenges.

12.9 Conclusions.

12.10 References.

13. MALDI-FTMS of Whole Cell Bacteria (Jeffrey J. Jones, MichaelJ. Stump, and Charles L. Wilkins).

13.1. Introduction.

13.2. Fundamentals of MALDI-FTMS.

13.3. Fundmentals of Complex Biological Analysis.

13.4 Whole Cell Characterization Through MALDI-FTMS.

13.5 Recombinant Over-Expressed Proteins Desorbed Directly fromWhole Cells.

13.6. Conclusions.

13.7. References.

14. A Review of Antibody Capture and Bacteriophage Amplificationin Connection with the Direct Analysis of Whole Cell Bacteria byMALDI-TOF-MS (Kent J. Voorhees and Jon C. Rees).

14.1. Introduction.

14.2. Bacterial Identification.

14.3. Immunocapture of Bacterial Mixtures.

14.4. Bacteriophage Amplification of Bacteria.

14.5. Conclusions.

14.6. References.

15. Discrimination and Identification of Microorganisms byPpyrolysis Mass Spectrometry: From Burning Ambitions to CoolingEmbers - A Historical Perspective (Éadaoin Timmins and RoystonGoodacre).

15.1 Introduction to microbial characterisation.

15.2 Principles of PyMS.

15.3 Early developments and investigations (1952 to 1985).

15.4 The mid 1980s and beyond.

15.5 The move from cluster analyses to neural networks.

15.6 Reproducibility of PyMS.

15.7 Acknowledgements.

15.8 References.


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