Nano-etched fused-silica capillary used for on-line preconcentration and electrophoretic separation of bacteriophages from large blood sample volumes with off-line MALDI-TOF mass spectrometry identification

Varování

Publikace nespadá pod Filozofickou fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
Autoři

HORKÁ Marie KARÁSEK Pavel ŠALPLACHTA Jiří RŮŽIČKA Filip ŠTVERÁKOVÁ Dana PANTŮČEK Roman ROTH Michal

Rok publikování 2020
Druh Článek v odborném periodiku
Časopis / Zdroj Microchimica Acta
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www https://link.springer.com/article/10.1007/s00604-020-4154-6
Doi http://dx.doi.org/10.1007/s00604-020-4154-6
Klíčová slova Capillary electrophoresis; MALDI-TOF mass spectrometry; Nano-etched fused-silica capillary; Phage propagation; Staphylococcal bacteriophages; Supercritical water
Přiložené soubory
Popis The properties of staphylococcal phages from the Siphoviridae, Podoviridae, and Myoviridae families were monitored using capillary electrophoretic methods on fused-silica capillaries with different morphology of surface roughness. Isoelectric points of the examined phages were determined by capillary isoelectric focusing in the original, smooth fused-silica capillary, and they ranged from 3.30 to 3.85. For capillary electrophoresis of phages, fused-silica capillaries with the "pock" and "cone" roughened surface types were prepared by etching a part of the capillary with supercritical water. The best resolution of the individual phages (to range from 3.2 to 4.6) was achieved with the "cone" surface-type fused-silica capillary. Direct application of phage K1/420 at the infection site, represented by human plasma or full blood spiked with Staphylococcus aureus, was on-line monitored by micellar electrokinetic chromatography. The phage particles were dynamically adhered onto the roughened surface of the capillary from 10 mu L of the prepared sample at the optimized flow rate of 6.5 mu L min(-1). The limit of detection was determined to be 10(4) phage particles. The linearity of the calibration lines was characterized by the regression coefficient, R-2 = 0.998. The relative standard deviation (RSD) of the peak area, calculated from ten independent measurements, was (+/-) 2%. After analysis, viability of the detected phages was verified by the modified "double-layer drop assay" method, and collected phage fractions were simultaneously off-line analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Graphical abstract
Související projekty:

Používáte starou verzi internetového prohlížeče. Doporučujeme aktualizovat Váš prohlížeč na nejnovější verzi.