Analysis of Bacillus globigii spores by CE.
It is imperative in today's world that harmful airborne or solution-based microbes can be detected quickly and efficiently. Bacillus globigii (Bg) spores are used as a simulant for Bacillus anthracis (Ba) due to their similar shape, size, and cellular makeup. The utility of CE to separate and detect low levels of Bg spore concentrations will be evaluated. To differentiate spores from background particulates, several dyes, including fluorescamine, C-10, NN-127, Red-1c, and indocyanine green (ICG), were utilized as noncovalent labels for proteins on the Bg spore surface, as well as for HSA and homoserine standards. On-column labeling, with dye present in the running buffer, was utilized to obtain greater sensitivity and better separation. CE with LIF detection enables interactions between the dye and spore surface proteins to be observed, with enhanced fluorescence occurring upon binding of the dye to surface protein. Resulting electropherograms showed unique fingerprints for each dye with Bg spores. Migration times were under 10 min for all dye-spore complexes, with net mobilities ranging from 3.5x10(-4) to 6.9x10(-4) cm(2) V(-1) s(-1), and calibration curves yielded correlation coefficients of 0.98 or better for four of the dyes studied.
Chichester
Kimberly D
KD
Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA.
Silcott
David B
DB
Colyer
Christa L
CL
D001407:Bacillus / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry; D004396:Coloring Agents; D003503:Cyclobutanes; D019075:Electrophoresis, Capillary / Q000379:methods*; D005450:Fluorescamine; D005453:Fluorescence; D005456:Fluorescent Dyes; D007208:Indocyanine Green; D007211:Indoles; D011485:Protein Binding; D013171:Spores, Bacterial / Q000302:isolation & purification*
D001426:Bacterial Proteins; D004396:Coloring Agents; D003503:Cyclobutanes; D005456:Fluorescent Dyes; D007211:Indoles; C511351:Red-1c compound; D005450:Fluorescamine; D007208:Indocyanine Green