|
|
|
|
| Institute: |
Lab or Branch |
| U. S. Army |
Chemical Biological Point Detection Team,
Aberdeen Proving Ground, MD |
|
| Title: |
| Effect Of Harvest Time On The LC-MS Profile
Of Vegetative And Sporulated Bacteria Using An In-House
Biological Sample Processing System |
|
| Authors: |
| R.E. Jabbour, S.V. Deshpande, W.M. Maswadeh,
A.P. Snyder |
|
| Abstract: |
| Characterization of microorganisms is being
studied with various bioanalytical tools. Many factors
must be considered in establishing genome or proteome
databases. One factor that affects pathogen characterization
is the growth phase. The technique of liquid chromatography-mass
spectrometry (LC-MS) of bacterial protein extracts harvested
at various growth phases is being investigated for microorganism
differentiation utility. This differentiation process
is achieved at each growth phase by determining useful
protein biomarkers of the bacterial extracts. Bacterial
samples are processed using an in-house biological sample
processing system (BSPS). The BSPS is being developed
to provide a rugged, rapid and reproducible procedure
capable of protein profiling of various pathogens. The
BSPS consists of three sequentially compatible compartments
for protein isolation and purification, coupled to micro-
or nano- liquid chromatography, which is interfaced to
an electrospray (ESI)-ion trap MS system. Following the
BSPS-MS analysis, the collected data are processed by
software that integrates the LC-MS peaks with subsequent
mass deconvolution. These masses are archived in an in-house
database and are analyzed with multivariate data analysis
for detection and differentiation purposes. This presentation
will address the effect of harvest time on the LC-MS profile
of bacteria in vegetative and sporulated states. The impact
of growth time on the in-house bacterial proteome database
and the reproducibility of the BSPS-MS analyses will be
presented. |
|
| |
|
