Fused silica packed microcolumns as micropreparative tools in protein analytical studies

Flurer, Cheryl L.; Borra, Claudio; Beale, Stephen C.; Novotný, Miloš V.

doi:10.1021/ac00168a036

Cited by 28 publications

(13 citation statements)

References 30 publications

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“…We believe that with improved transfer procedures and detection conditions even lower detection limits should be feasible. We have demonstrated previously the use of BQCA as a fluorogenic reagent (8)(9)(10). The detection limits for the BNA derivatives shown here are in the range of the detection limits for BQCA derivatives.…”

Section: Resultsmentioning

confidence: 59%

3‐Benzoyl‐2‐naphthaldehyde, a new fluorogenic reagent for microcolumn liquid chromatography–laser‐induced fluorescence detection of amino acids

Hsieh

Beale

Wiesler

et al. 1989

J Microcolumn Separations

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Abstract. 3-Benwyl-2-naphthaldehyde (BNA) was synthesized and characterized as a precolumn derivatization reagent for fluorescence detection. The detection limits for standard amino acids derivatized with BNA are in the femtogram range. The optimum reaction conditions for derivatization and spectral features of BNA-derivatized amino acids was investigated. The standard amino acid derivatives were adequately resolved by microcolumn liquid chromatography. A minute quantity (646 fmol) of trypsin was hydrolyzed to demonstrate analysis by microcolumn liquid chromatography with laser-induced fluorescence detection.

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Section: Resultsmentioning

confidence: 59%

3‐Benzoyl‐2‐naphthaldehyde, a new fluorogenic reagent for microcolumn liquid chromatography–laser‐induced fluorescence detection of amino acids

Hsieh

Beale

Wiesler

et al. 1989

J Microcolumn Separations

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show abstract

“…It is thus logical that a sensitive biomolecule can greatly benefit from the use of microcolumns having drastically reduced amounts of sorptive materials. Recent investigations by our laboratory emphasize the superior inertness of slurry-packed fused-silica microcolumns with proteins (14,15). As shown in Figure 3, which isan example of size exclusion chromatography, elution of several standard proteins was accomplished at the level of roughly 200 pg each (14).…”

Section: Microcolumn Separations Of Proteins and Their Degradation Prmentioning

confidence: 99%

“…Recent investigations by our laboratory emphasize the superior inertness of slurry-packed fused-silica microcolumns with proteins (14,15). As shown in Figure 3, which isan example of size exclusion chromatography, elution of several standard proteins was accomplished at the level of roughly 200 pg each (14). Here, another important advantage of microcolumns -that cf the increased mass detection sensitivity (1 6,17) -becomes essential.…”

Section: Microcolumn Separations Of Proteins and Their Degradation Prmentioning

confidence: 99%

“…At this date, we have successfully demonstrated the preparation of fused-silica microcolumns for reversed-phase and size exclusion modes (14,15). Microcolumns packed with other sorption materials are highly desirable for their additional selectivities.…”

Section: Microcolumn Separations Of Proteins and Their Degradation Prmentioning

confidence: 99%

“…Consequently, microcolumns do not provide greater solute resolution, in this capacity, when compared with conventional columns. Their most valuable asset is their remarkable inertness, as demonstrated for model proteins through the signal vs. solute concentration plots (14).…”

Section: Microcolumn Separations Of Proteins and Their Degradation Prmentioning

confidence: 99%

See 2 more Smart Citations

Recent advances in the isolation and structural studies of biomacromolecules using microcolumn techniques

Novotný

1990

J Microcolumn Separations

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Abstract. Microcolumn liquid chromatography and capillary electrophoresis facilitate isolation of biological macromolecules at subnanogram levels. In order to characterize structurally such small amounts of labile materials, it is essential to develop new nanoscale sample treatment approaches and highly sensitive analytical techniques for the determination of biomolecular fragments. Progress in this area is summarized, using selected examples of the amino acid, peptide, and oligonucleotide separations.

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Mass spectrometry and the age of the proteome

1998

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Mass spectrometry has become an important technique to correlate proteins to their genes. This has been achieved, in part, by improvements in ionization and mass analysis techniques concurrently with large-scale DNA sequencing of whole genomes. Genome sequence information has provided a convenient and powerful resource for protein identiÐcation using data produced by matrix-assisted laser desorption/ionization time-of-Ñight (MALDI/TOF) and tandem mass spectrometers. Both of these approaches have been applied to the identiÐcation of electrophoretically separated protein mixtures. New methods for the direct identiÐcation of proteins in mixtures using a combination of enzymatic proteolysis, liquid chromatographic separation, tandem mass spectrometry and computer algorithms which match peptide tandem mass spectra to sequences in the database are also emerging. This tutorial review describes the principles of ionization and mass analysis for peptide and protein analysis and then focuses on current methods employing MALDI and electrospray ionization for protein identiÐcation and sequencing. Database searching approaches to identify proteins using data produced by MALDI/TOF and tandem mass spectrometry are also discussed.

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Fused silica packed microcolumns as micropreparative tools in protein analytical studies

Cited by 28 publications

References 30 publications

3‐Benzoyl‐2‐naphthaldehyde, a new fluorogenic reagent for microcolumn liquid chromatography–laser‐induced fluorescence detection of amino acids

3‐Benzoyl‐2‐naphthaldehyde, a new fluorogenic reagent for microcolumn liquid chromatography–laser‐induced fluorescence detection of amino acids

Recent advances in the isolation and structural studies of biomacromolecules using microcolumn techniques

Mass spectrometry and the age of the proteome

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