We propose to design, build and develop a compact tandem time-of-flight mass spectrometer for high throughput proteomics research based upon several technologies being developed in our laboratory. In particular the first mass analyzer (MSI) will utilize mass-correlated acceleration to focus ions with different kinetic energies and independently of mass to a focus point that is the entrance to the second mass analyzer (MS2). MS2 will be an endcap reflectron, which provides high order (nearly quadratic) kinetic energy focusing. The collision chamber will be located early on in the flight tube of MS1 as it is an essential feature of this instrument that there are no decelerating or re-accelerating fields, either constant or pulsed, between the ion source and the entrance to the endcap reflectron. Thus, fragment ions associated with a mass-selected precursor ion, and produced by collision-induced dissociation (CID) or by metastable decomposition, will be recorded at the same flight time. The high energy bandwidth endcap reflectron does not require stepping or scanning to record the entire product ion mass range in each time-of-flight cycle. In addition, because this reflectron also does not require reacceleration of after collision, it will be possible to utilize the full ion kinetic energy (10-20 keV) in collisional activation. The instrument will be utilized in a number of ongoing research projects involving peptidic and glycolipid antigens, the structures of lipid A, and protein arrays on a chip. In addition, a novel planar endcap reflectron design will be developed that is compatible with orthogonal acceleration. This will be used on an existing electron impact/orthogonal acceleration instrument, and will be further developed using atmospheric pressure MALDI as the ionization source. This technology is expected to bring high performance and tandem capabilities to a simple, compact instrument that can be utilized in proteomics research.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM064402-04
Application #
6848330
Study Section
Special Emphasis Panel (ZRG1-BECM (01))
Program Officer
Edmonds, Charles G
Project Start
2002-02-01
Project End
2006-07-31
Budget Start
2005-02-01
Budget End
2006-07-31
Support Year
4
Fiscal Year
2005
Total Cost
$264,012
Indirect Cost
Name
Johns Hopkins University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Cotter, Robert J (2013) High energy collisions on tandem time-of-flight mass spectrometers. J Am Soc Mass Spectrom 24:657-74
Cotter, Robert J; Griffith, Wendell; Jelinek, Christine (2007) Tandem time-of-flight (TOF/TOF) mass spectrometry and the curved-field reflectron. J Chromatogr B Analyt Technol Biomed Life Sci 855:2-13
Irani, David N; Anderson, Caroline; Gundry, Rebekah et al. (2006) Cleavage of cystatin C in the cerebrospinal fluid of patients with multiple sclerosis. Ann Neurol 59:237-47
Reynolds, C Michael; Ribeiro, Anthony A; McGrath, Sara C et al. (2006) An outer membrane enzyme encoded by Salmonella typhimurium lpxR that removes the 3'-acyloxyacyl moiety of lipid A. J Biol Chem 281:21974-87
Gibbons, Henry S; Kalb, Suzanne R; Cotter, Robert J et al. (2005) Role of Mg2+ and pH in the modification of Salmonella lipid A after endocytosis by macrophage tumour cells. Mol Microbiol 55:425-40
Reynolds, C Michael; Kalb, Suzanne R; Cotter, Robert J et al. (2005) A phosphoethanolamine transferase specific for the outer 3-deoxy-D-manno-octulosonic acid residue of Escherichia coli lipopolysaccharide. Identification of the eptB gene and Ca2+ hypersensitivity of an eptB deletion mutant. J Biol Chem 280:21202-11
Gundry, Rebekah L; Edward, Roy; Kole, Thomas P et al. (2005) Disposable hydrophobic surface on MALDI targets for enhancing MS and MS/MS data of peptides. Anal Chem 77:6609-17
Wang, Dongxia; Thompson, Paul; Cole, Philip A et al. (2005) Structural analysis of a highly acetylated protein using a curved-field reflectron mass spectrometer. Proteomics 5:2288-96
Wooden, Stacey L; Kalb, Suzanne R; Cotter, Robert J et al. (2005) Cutting edge: HLA-E binds a peptide derived from the ATP-binding cassette transporter multidrug resistance-associated protein 7 and inhibits NK cell-mediated lysis. J Immunol 175:1383-7
Von Seggern, Christopher E; Gardner, Ben D; Cotter, Robert J (2004) Infrared atmospheric pressure MALDI ion trap mass spectrometry of frozen samples using a Peltier-cooled sample stage. Anal Chem 76:5887-93

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