Affinity monolith preconcentrators for polymer microchip capillary electrophoresis.
Developments in biology are increasing demands for rapid, inexpensive, and sensitive biomolecular analysis. In this study, polymer microdevices with monolithic columns and electrophoretic channels were used for biological separations. Glycidyl methacrylate-co-ethylene dimethacrylate monolithic columns were formed within poly(methyl methacrylate) microchannels by in situ photopolymerization. Flow experiments in these columns demonstrated retention and then elution of amino acids under conditions optimized for sample preconcentration. To enhance analyte selectivity, antibodies were immobilized on monoliths, and subsequent lysozyme treatment blocked nonspecific adsorption. The enrichment capability and selectivity of these affinity monoliths were evaluated by purifying fluorescently tagged amino acids from a mixture containing green fluorescent protein (GFP). Twenty-fold enrichment and 91% recovery were achieved for the labeled amino acids, with a >25 000-fold reduction in GFP concentration, as indicated by microchip electrophoresis analysis. These devices should provide a simple, inexpensive, and effective platform for trace analysis in complex biological samples.
Yang
Weichun
W
Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
Sun
Xiuhua
X
Pan
Tao
T
Woolley
Adam T
AT
D000327:Adsorption; D000596:Amino Acids / Q000032:analysis*; D000906:Antibodies / Q000276:immunology; D019075:Electrophoresis, Capillary / Q000379:methods; D046209:Electrophoresis, Microchip / Q000379:methods; D049452:Green Fluorescent Proteins / Q000737:chemistry; D009113:Muramidase / Q000276:immunology; D019904:Polymethyl Methacrylate / Q000737:chemistry*
D000596:Amino Acids; D000906:Antibodies; D049452:Green Fluorescent Proteins; D019904:Polymethyl Methacrylate; D009113:Muramidase