Electrophoretically mediated microanalysis for in-capillaryelectrical cell lysis and fast enzyme quantification by capillary electrophoresis

摘要: In this study, a novel capillary electrophoresis (CE)-based enzymatic assay was developed to evaluate activity in whole cells. β-Galactosidase expression used as an example, it is biomarker for assessing replicative senescence mammalian It catalyzes the hydrolysis of para-nitrophenyl-β-d-galactopyranoside (PNPG) into para-nitrophenol (PNP). The CE-based consisted four main steps: (1) hydrodynamic injection intact cells capillary, (2) in-capillary lysis these by using pulses electric field (electroporation), (3) PNPG β-galactosidase—released from lysed cells—by electrophoretically mediated microanalysis (EMMA) approach, and (4) on-line detection quantification PNP formed. method applied Escherichia coli well human keratinocyte at different stages. Results obtained CE were excellent agreement with those off-line cell lysates which proves efficiency approach developed. This work shows first time that membranes can be disrupted electroporation released enzyme subsequently quantified same capillary. Enzyme after their has never been conducted CE. automated, economic, eco-friendly, simple conduct. attractive applications bacteria or early disease diagnostics insights development biology.