Amplification of lysozyme signal detected in capillary electrophoresis using mixed polymer brushes coating with switchable properties

Abstract

In this work, a mixed polymer brushes based on poly(2-methyl-2-oxazoline) (PMOXA) and poly(acrylic acid) (PAA) coated capillary with switchable protein adsorption/desorption properties was developed and applied for on-line extraction and preconcentration of lysozyme. The study of electroosmotic flow (EOF) and fluorescence microscope showed that the inner surface charge of PMOXA/PAA mixed brush coated capillary displayed the switchable behavior toward the change of pH value and ionic strength (I), and PMOXA/PAA mixed brushes coated capillary could adsorb high amounts of lysozyme at pH 7 (I = 10−5 M), and the most of adsorbed lysozyme could then be desorbed at pH 3 (I = 10−1 M). Subsequently, this coated capillary with switchable lysozyme adsorption/desorption ability was applied for on-line extraction and preconcentration of lysozyme during capillary electrophoresis (CE) performance. Under the process of on-line preconcentration, the detection signal (peak area) of lysozyme obtained in PMOXA/PAA coated capillary was 26 times that obtained in bare capillary under normal CE while the contour chain length of PAA was 1.56 times that of PMOXA. Moreover, the value of low detection limit (LOD) of lysozyme using above coated capillary under on-line preconcentration method reached to 4.5 × 10−9 mg/mL, and 1 × 105-fold sensitivity enhancement was realized for lysozyme as compared with the bare capillary under normal CE.