Bacterial characterization using protein profiling in a microchip separations platform.
A rapid microanalytical protein-based approach to bacterial characterization is presented. Chip gel electrophoresis (CGE) coupled with LIF detection was used to analyze lysates from different bacterial cell lines to obtain signature profiles of the soluble protein composition. The study includes Escherichia coli, Bacillus subtilis, and Bacillus anthracis (Delta Sterne strain) vegetative cells as well as endospores formed from the latter two species as model organisms to demonstrate the method. A unified protein preparation protocol was developed for both cell types to streamline the benchtop process and aid future automation. Cells and spores were lysed and proteins solubilized using a combination of thermal and chemical lysis methods. Reducing agents, necessary to solubilize spore proteins, were eliminated using a small-scale rapid size-exclusion chromatography step to eliminate interference with down-stream protein labeling. This approach was found to be compatible with nonspore cells (i.e., vegetative cells) as well, not adversely impacting the protein signatures. Data are presented demonstrating distinct CGE protein signatures for our model organisms, suggesting the potential for discrimination of organisms on the basis of empirical protein patterns. The goal of this work is to develop a fast and field-portable method for characterizing bacteria via their proteomes.
Pizarro
Shelly A
SA
Sandia National Laboratories, Biosystems Research Department, Livermore, CA 94551-9292, USA.
Lane
Pamela
P
Lane
Todd W
TW
Cruz
Evelyn
E
Haroldsen
Brent
B
VanderNoot
Victoria A
VA
D001419:Bacteria / Q000145:classification* / Q000378:metabolism; D001426:Bacterial Proteins / Q000302:isolation & purification*; D046209:Electrophoresis, Microchip / Q000379:methods*
D001426:Bacterial Proteins