Abstract
Development of capillary tubes internally doped with enzymes is of great interest for microfluidic reactions, and kinetic doping could provide a facile, inexpensive method for their manufacture. Kinetic doping has previously been demonstrated to have a high loading capacity with thin films coated on flat-surface coverslips. Dip coating of these surfaces was developed with the eventual intention to coat different shapes and sizes of substrates. In this study, we expanded the use of kinetic doping to internally-coated capillary tubes. Parameters for internally doping capillary tubes were developed with rhodamine 6G, which produced internally-coated thin films with a 90 nm thickness. Horseradish peroxidase (HRP) was then loaded into the capillary tubes, with a 47,000× increase in concentration over the loading solution. After excluding surface-adsorbed protein, the increase in HRP concentration in the thin films over the loading solution was determined to be 9850×. The activity of the loaded HRP was determined to be 0.019 ± 0.003 U/mg and shown to have a stronger resistance to denaturation by methanol.
Highlights
IntroductionMany researchers have investigated the encapsulation of enzymes in silica in order to gain the benefits of immobilization, namely, high reusability and higher chemical and thermal stability [1,2,3,4,5,6,7,8,9,10,11]
Many researchers have investigated the encapsulation of enzymes in silica in order to gain the benefits of immobilization, namely, high reusability and higher chemical and thermal stability [1,2,3,4,5,6,7,8,9,10,11].Sol-gel-based technologies are especially promising, with their ease of synthesis and high loading capacity [9,12]
We extended the kinetic doping technique developed by our group to internally coat capillary tubes with silica thin films with entrapped horseradish peroxidase (HRP)
Summary
Many researchers have investigated the encapsulation of enzymes in silica in order to gain the benefits of immobilization, namely, high reusability and higher chemical and thermal stability [1,2,3,4,5,6,7,8,9,10,11]. Chemical modification of the surfaces for immobilization of enzymes in devices is commonly required [15,16,17,18], increasing the difficulty and expense of production Overcoming these barriers to enable inexpensive and facile manufacturing of glass capillary tubes with high concentrations of immobilized enzymes could be of great benefit. Kinetic doping has been shown to exhibit a high dopant capacity, produce films with good retention of enzyme activity, and nearly instantaneous response time [33] This could lead to an inexpensive manufacturing process for enzyme-coated glass capillary tubes that is low on both material and labor cost, which is significant for potential commercialization. This was the first time that an entrapped enzyme had been internally coated onto a capillary tube using the sol-gel method
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