Silica nanoparticles are most commonly modified with amino-silanes, followed by post-modification activation for protein immobilization. In this work, epoxy-functionalized silica nanoparticles were prepared by modification with glycidyloxypropyl trimethoxysilane (GPTMS) for direct protein immobilization. Silica nanoparticles possessed an average size of 46 nm, but increased to 63 nm after GPTMS modification. Reaction time, reaction temperature and GPTMS content had no significant effect on particle size. Zeta potential of SiO 2 changed from −26mV to +38mV after modification. Fourier-transformed infrared spectroscopy revealed alkyl C-H bending and stretching bands at 2944 cm −1 , 1343 cm −1 and 1465 cm −1 , respectively, for the modified nanoparticles. Fluorescein cadaverine was found to bind to GPTMS-modified SiO 2 , but not to bare SiO 2 , indicating the chemical reactivity of epoxy groups on the modified nanoparticle with amines. Finally, fluorescently labeled bovine serum albumin (BSA) was used as a model protein to investigate the capacity of epoxy-SiO 2 nanoparticles for protein immobilization. The results showed that more proteins were immobilized on the particle with longer reaction time, higher NaCl concentration, lower pH, and less GPTMS content. More importantly, proteins bound to epoxy-SiO 2 nanoparticle were highly stable. Under optimized reaction conditions, as much as 25 mg BSA/g nanoparticle was covalently attached to the nanoparticle. The epoxy silane modification of silica nanoparticles offers a reactive surface for one-step and high-density protein immobilization.
epoxysilane, silica nanoparticle, protein immobilizationIn the past few years, nanomaterials of various shapes (particles, wires, rods, tubes, etc.) have been synthesized and characterized. They have been found to possess unique size-dependent properties in terms of quantum effect, specific surface area, electric conductivity, mechanical strength, and catalytic reactivity [1] . Consequently, nanomaterials have been found wide applications in the field of biological analysis and detection, biological imaging and drug delivery [2,3] . Among them, silica nanoparticles have been studied extensively due to their easy synthesis, readily available surface modification, good dispersibility and stability in water, and biocompatibility [4] . Mesoporous silica nanoparticles can be employed as drug delivery vehicles for controlled release [5] . Surface-coated silica nanoparticles act as a carrier for multiple optical [6,7] and electrochemical [8] labels to enhance detection signal. Dye-doped silica nanoparticles are more resistant to photo-bleaching than fluorescent dye molecules and much less toxic than quantum dots. They therefore offer distinct advantages when applied to in vivo fluorescence imaging [9,10] . Ruthenium tris(bipyridyl)-doped silica nanoparticles [11][12][13] have been used as electrochemiluminescent labels to provide large quantities of Ru(bpy) 3 2+ molecules for signal detection. Furthermore, silica coating on other