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Basile Lab

Basile Research Group

Franco Basile, Ph.D., Principal Investigator

Dept 3838

Department of Chemistry

Department of Chemistry

University of Wyoming

1000 E. University Ave.

Office (307) 766-4376

Laboratory (307) 766-2991

Laramie, WY 82071

Email: basile@uwyo.edu

Basike Research Group

Basile Lab

Our Research

" ...the most difficult portion of the analytical process is that involving isolation of the desired constituent in a measurable state. The actual process of measurement is simple."

Ernest B. Sandell and Phillip J. Elving, Methods of Analytical Chemistry, Chapter 1 (p. 7) in Treatise on Analytical Chemistry Part I Theory and Practice; I. M. Kolthoff and Phillip J. Elving (Eds.), 1959

Funding for this research has been provided by:

National Science FoundationEPSCOR Wyoming logoWyoming INBRENational Institutes of Health
University of Wyoming School of Natural ResourcesDefense Threat Reduction Agency

Analytical and Bioanalytical Chemistry/Mass Spectrometry

Protein Thermal Decomposition/Digestion (TDD): site-specific protein digestion at Asp and Cys in 10 sec

Our laboratory developed a technique for the rapid (10-30 sec) digestion of proteins based on their controlled thermal degradation.  Our approach is able to perform site-specific cleavages at both Asp and Cys with minimum side reactions and modifications.  This Thermal Decomposition/Digestion (TDD) process can be performed on solid samples and current efforts are focusing on the implementation of this technique for the analysis of biological tissue samples (i.e., imaging MALDI-MS/MS).

Read our articles in J. Am. Soc. Mass Spectrom. (2011) 22: 1926-1940 and J. Anal. Appl. Pyrolysis (2017)

Imaging MALDI-MS and Thermal Decomposition/Digestion (TDD): towards on-tissue digestion and on-tissue protein identification

Proof-of-principle imaging MALDI-MS of the TDD products of lysozyme: Detected peptides are from site-specific cleavages at Asp and Cys.  Images of the spatial distribution of these peptide products are shown below in a proof-of-principle experiment demonstrating applicability of this approach for on-tissue protein id and tissue imaging by MALDI-MS. The solvent-free nature of the TDD process prevents product delocalization, as is the case in the standard protocol using on-tissue trypsin digest. Read our article in Analytical Chem. (2017).

Imaging results

Metabolomics/Lipidomics

Our laboratory implements several MS techniques (GC-MS, GC-MS/MS, and LC offline MALDI-MS/MS) for the analysis of lipids, carbohydrates and metabolites in microorganisms, bees and soil.  Recent work from our laboratory identified several hydroxy-fatty acids (some believed to be associated with lipid A) and polyunsaturated fatty acids in the microorganism Gemmata obscuriglobus. (Project in collaboration with Dr. Naomi Ward; Read the article in J. Bacteriology, 2016, 198, 221-236 journal link).

Image of metabolomics/lipidomics results

Increasing the reproducibility of intact bacteria MALDI-MS profiling

The goal of this project is the development of a safe, automated and highly reproducible technique to prepare microbial samples for MALDI-MS profiling.  By implementing a spray-deposition method, we have identified that the largest source of variation comes from the surface heterogeneity of the MALDI matrix.

Read the article in Analytical Chem. 2012, 84, 9971-9980

Image of MALDI-MS profiling results

 

Developing New Tools for High Throughput Proteomics using Non-Enzymatic Protein Digestion Methods:

Online Microwave D-Cleavage: cleavage at aspartic acid and disulfide bonds

Microwave D-cleavage: Our laboratory pioneered the online microwave-heated site-specific acid hydrolysis at aspartic acid (C-terminus) and disulfide bond cleavage in proteins.  The microwave D-cleavage technique allows the simultaneous digestion at aspartic acid and disulfide bonds in 5 minutes and couples online with LC-ESI-MS/MS for data-dependent analysis.  This technique successfully detected and identified all proteins tested as well as proteins in an E. coli cell lysate at a high confidence level. Read the article in J. Proteome Res. (2008) 7: 1012-1026

Image of Microwav D-Cleavage process

We are currently focusing on the development of a fully integrated sample preparation technique for proteomics (a.k.a., proteomic reactor) combining microwave D-cleavage with electrochem WY-cleavage to digest proteins online in under 6 min.  The resulting peptides are well suited for CID MS/MS analysis since they average about 10 amino acids in length (tryptic peptides average about 9 amino acids in length). Read the article in Analytical Chem. (2011) 83: 35

Other projects currently under way in our laboratory:

Renewable Energy Resources and Environmental MS

  • Broad spectrum analysis of coal samples: rapid extraction methods; GC-MS characterization and analysis, Read article in Energy & Fuels (2014)

Image of Microwave heating process

Metabolomics

  • Lipidomics: GC-MS and LC-offline MALDI-MS/MS
  • instrumentation development: continuous-flow liquid microsampling for surface analysis

 

Research Collaborators

Erika Belmont, Dept. of Mechanical Eng., University of Wyoming

Wei Guo, Dept. of Animal Sciences, University of Wyoming

Naomi Ward, Molecular Biology, University of Wyoming

Katie Li, Dept. of Chemical Eng., University of Wyoming

Linda van Diepen, Dept. of Ecosystem Science and Management, University of Wyoming

Michael Dillon, Dept. Zoology & Physiology/Program in Ecology, University of Wyoming

Debashis Dutta,  Dept. of Chemistry, University of Wyoming

Scott McLuckey, Dept. of Chemistry, Purdue University

Jessica Prenni, Colorado State University

John Belisle, Colorado State University

Claudia Molins, Centers for Disease Control and Prevention (Ft. Collins, CO)

Eva Ferre-Pikal,  Dept. of Electrical Eng., University of Wyoming

Vladimir Alvarado,  Dept. of Chemical Eng., University of Wyoming

John Oakey, Dept. of Chemical Eng., University of Wyoming

David Bagley,  Dept. of Chemical Eng., University of Wyoming

Michael Urynowicz,  Dept. of Environmental Sciences, University of Wyoming

Junko Marr,  Dept. of Environmental Sciences, Colorado School of Mines

Kent Voorhees, Dept. of Chemistry, Colorado School of Mines

Donald L. Jarvis, Molecular Biology, University of Wyoming

Mark Gomelsky, Molecular Biology, University of Wyoming

Ami Wangeline, LCCC, Cheyenne, WY

Gregory Brown, Botany Dept., University of Wyoming

Steve Wolf, Statistics Dept., University of Wyoming


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Basile Lab

Basile Research Group

Franco Basile, Ph.D., Principal Investigator

Dept 3838

Department of Chemistry

Department of Chemistry

University of Wyoming

1000 E. University Ave.

Office (307) 766-4376

Laboratory (307) 766-2991

Laramie, WY 82071

Email: basile@uwyo.edu

Basike Research Group

1000 E. University Ave. Laramie, WY 82071
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