Microchip electrophoresis of N-glycans on serpentine separation channels with asymmetrically tapered turns.
We designed and fabricated microfluidic devices with serpentine separation channels and asymmetrically tapered turns, thus allowing high efficiency separations and minimizing band broadening associated with the "racetrack" effect. We evaluated the performance of these devices by measuring the variation in separation efficiency with separation length, electric field strength, taper ratio of the turns, and number of turns. N-Glycans derived from ribonuclease B and labeled with 8-aminopyrene-1,3,6-trisulfonic acid were electrophoretically separated on serpentine channels with separation lengths of 11, 18, 22, and 36 cm at electric field strengths from 750 to 1750 V/cm. Separations on the 36-cm channel produced plate numbers up to 940,000 with an analysis time under 3.1 min, whereas separations on the 22-cm channel had a shorter analysis time (less than 1.25 min), still with respectable efficiencies (up to 600,000 plates). Turn-induced dispersion was minimized with taper ratios 2 and 3, whereas having two or four 180° turns along with the separation length did not impact the overall efficiency. The developed device was used to analyze native and desialylated N-glycans derived from the blood serum of an ovarian cancer patient and a disease-free individual. Separation efficiencies similar to that achieved with the model glycans from ribonuclease B were attained for these biological samples.
Department of Chemistry, Indiana University, Bloomington, Indiana, USA.
Zexi
Z
Zhuang
Indranil
I
Mitra
Ahmed
A
Hussein
Milos V
MV
Novotny
Yehia
Y
Mechref
Stephen C
SC
Jacobson
D046209:Electrophoresis, Microchip / Q000379:methods*; D006801:Humans; D007536:Isomerism; D046210:Microfluidic Analytical Techniques; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D011721:Pyrenes / Q000737:chemistry; D012260:Ribonucleases / Q000378:metabolism; D046948:Secologanin Tryptamine Alkaloids
C103430:8-aminopyrene-1,3,6-trisulfonic acid; D011134:Polysaccharides; D011721:Pyrenes; D046948:Secologanin Tryptamine Alkaloids; C009244:serpentine (alkaloid); D012260:Ribonucleases; C029600:ribonuclease B
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