Microbore high-performance liquid chromatography—mass spectrometry for the analysis of proteolytic digests by continuous-flow fast-atom bombardment mass spectrometry

Caprioli, Richard M.; Moore, William T.; DaGue, Beverly B.; Martin, M. Katie

doi:10.1016/s0021-9673(00)94807-6

Cited by 46 publications

(14 citation statements)

References 8 publications

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“…HPLC column flow rates in the range of 5 ,uL/min are also compatible with continuous-flow fast atom bombardment MS, and the use of CF-FAB for LC/MS peptide mapping experiments at sensitivities comparable to the ones reported here have been demonstrated (Caprioli et al, 1988;Henzel et al, 1990). Hemling and coworkers (1990) have pointed out several advantages of electrospray ionization compared to CF-FAB with respect to ease of operation, significantly extended mass range due to multiple charging, and generality, including detection of glycopeptides.…”

Section: Discussionmentioning

confidence: 65%

Analytical and micropreparative peptide mapping by high performance liquid chromatography/electrospray mass spectrometry of proteins purified by gel electrophoresis

et al. 1993

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We report the use of microbore reverse-phase high performance liquid chromatography connected on-line to an electrospray mass spectrometer for the separation/detection of peptides derived by proteolytic digestion of proteins separated by polyacrylamide gel electrophoresis. A small fraction (typically 10% of the total) of the peptides eluting from the column was diverted through a flow-splitting device into the ion source of the mass spectrometer, whereas the majority of the peptide samples was collected for further analyses. We demonstrate the feasibility of obtaining reproducible peptide maps from submicrogram amounts of protein applied to the gel and good correlation of the signal detected by the mass spectrometer with peptide detection by UV absorbance. Furthermore, independently verifiable peptide masses were determined from subpicomole amounts of peptides directed into the mass spectrometer.The method was used to analyze the 265-kDa and the 280-kDa isoforms of the enzyme acetyl-coA carboxylase isolated from rat liver. The results provide compelling evidence that the two enzyme isoforms are translation products of different genes and suggest that these approaches may be of general utility in the definitive comparison of protein isoforms. We furthermore illustrate that knowledge of peptide masses as determined by this technique provides a major advantage for error-free data interpretation in chemical high-sensitivity peptide sequence analysis.

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Section: Discussionmentioning

confidence: 65%

Analytical and micropreparative peptide mapping by high performance liquid chromatography/electrospray mass spectrometry of proteins purified by gel electrophoresis

et al. 1993

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“…The sensitivity of the Hb tryptic peptide analysis was improved 10 to 100 times which allowed pmol level studies by the use of microHPLC/FAB combination as already published by Caprioli et al (42). We achieved an on-line trypsin hydrolysis-mass spectrometry with a very small dead volume (50 pl) which allows a continuous separation by microLC of big peptides, tryptic cleavage, FA6 mapping and if necessary,…”

Section: Hb Mckees Rockmentioning

confidence: 97%

New Results of Hemoglobin Variant Structure Determinats by Fast Atom Bombardment Mass Spectrometry

et al. 1990

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Fast atom bombardment mass spectrometry has already been used for the identification of mutations in abnormal human hemoglobin chains. This paper presents new results obtained with this technique. The methodology used here is compared with more conventional biochemical techniques and automated microsequencing. In every case, a well-chosen combination of peptide-high performance liquid chromatography, mass spectrometry, amino acid analysis, and sequence analysis led rapidly to the identification of the mutant. The high sensitivity of these techniques holds great promise for the analysis of molecular abnormalities in various genetic disorders presently detectable only by the application of a molecular biological approach.

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“…Caprioli's group, then at Texas Medical School in Houston published their work on the continuous-flow FAB interface for direct coupling of microbore and slurry packed C 18 fused silica capillary columns. [42] Another topic that must be properly placed within the context of these studies on electrospray and new LC-MS interfaces is the parallel work that led to the development of 'Ion Spray', as reported in 1987 by Jack Henion, Tom Covey and Andries Bruins at Cornell University (see below), the latter on leave from Groningen, The Netherlands. These authors prompted the development of atmospheric pressure ionization (API) based on the earlier work on APCI/MS (see Part I) carried out in Houston by the group of Evan Horning and described in a December 1975 publication in Analytical Chemistry.…”

Section: The Arrival Of Modern Lc-msmentioning

confidence: 99%

From large analogical instruments to small digital black boxes: 40 years of progress in mass spectrometry and its role in proteomics. Part II 1985–2000

Gelpí

2009

J. Mass Spectrom.

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Microbore high-performance liquid chromatography—mass spectrometry for the analysis of proteolytic digests by continuous-flow fast-atom bombardment mass spectrometry

Cited by 46 publications

References 8 publications

Analytical and micropreparative peptide mapping by high performance liquid chromatography/electrospray mass spectrometry of proteins purified by gel electrophoresis

Analytical and micropreparative peptide mapping by high performance liquid chromatography/electrospray mass spectrometry of proteins purified by gel electrophoresis

New Results of Hemoglobin Variant Structure Determinats by Fast Atom Bombardment Mass Spectrometry

From large analogical instruments to small digital black boxes: 40 years of progress in mass spectrometry and its role in proteomics. Part II 1985–2000

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