Instrumentation strategy for investigation of bacteriophage 812 proteome by on-line combination of capillary electrophoresis, electrochromatography and liquid chromatography with mass spectrometry
| Authors | |
|---|---|
| Year of publication | 2005 |
| Type | Article in Proceedings |
| Conference | 18th International Symposium on MicroScale Bioseparations (MSB) |
| MU Faculty or unit | |
| Citation | |
| Web | http://www.casss.org/meetings/2005msb/default.htm |
| Field | Analytic chemistry |
| Keywords | BACTERIOPHAGE; PROTEOME; CAPILLARY ELECTROPHORESIS; ELECTROCHROMATOGRAPHY; LIQUID CHROMATOGRAPHY; MASS SPECTROMETRY |
| Description | We have developed a new miniaturized instrumentation for combination of the microscale liquid phase separation techniques with mass spectrometry (MS). Its capability will be demonstrated on analyses of proteins prepared from bacteriophage 812. The optimum conditions for growing bacteriophages, preparation of phage proteins and their tryptic digests, and the capillary separations provided a satisfactory amount of peptides for the protein identification by MS with electrospray. The capillary separation columns and nano-spray needles made of narrow bore (ID 5 50 micrometers) silica-fused capillaries were connected by an original laboratory-made interface. The ultimate goal of our research is to contribute to the understanding of the molecular principle of phage therapy. The alarming increase in antibiotic-resistant bacteria has renewed interest in bacteriophage therapy of various human infections. The knowledge of bacteriophages lytic enzymes and cell growth inhibitors can potentially lead to the development of low-molecular species with a specific therapeutic activity. |
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