Microflow-based automated chemistries: Application to protein sequencing Journal Article
| Authors: | Powell, M.; Tempst, P. |
| Article Title: | Microflow-based automated chemistries: Application to protein sequencing |
| Abstract: | We describe a microflow-based instrument, consisting of multiple rotary valves, capillary tubing, and miniaturized reaction vessels, for the purpose of performing automated chemical and biochemical reactions on a very small scale (i.e. submicroliter volumes). The novelty is that close to 100% of the reaction end products are available in a minimal volume (≤5 μL) inside a pressurized microvial for subsequent analysis. This makes the system compatible with capillary HPLC and, in principle, with continuous-flow nanoelectrospray mass spectrometry. Total control of flow path combinations and directions, temperatures and gas pressures enables precise execution of complex biochemical laboratory procedures. Instrument performance was convincingly demonstrated by partially sequencing 100 fmol of an intact protein using classical Edman chemistry in combination of capillary-bore liquid chomatography. To our knowledge, this is the smallest amount of protein ever reported to be successfully analyzed in this way. Additional application that merge enzymatic and chemical procedures with high-sensitivity analytical devices can be envisioned in the future. |
| Keywords: | mass spectrometry; protein analysis; protein degradation; protein; enzyme activity; chemistry; amino acid sequence; capillary electrophoresis; high performance liquid chromatography; chromatography, high pressure liquid; chemical reaction; sequence analysis, protein; indicators and reagents; article; lactoglobulins |
| Journal Title: | Analytical Chemistry |
| Volume: | 73 |
| Issue: | 4 |
| ISSN: | 0003-2700 |
| Publisher: | American Chemical Society |
| Date Published: | 2001-02-15 |
| Start Page: | 776 |
| End Page: | 786 |
| Language: | English |
| DOI: | 10.1021/ac000926p |
| PUBMED: | 11248892 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 21 May 2015 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work