Biomolecular screening with novel organosilica microspheres

Abstract

Organosilica microspheres synthesised via a novel surfactant-free emulsion-based method show applicability towards optical encoding, solid-phase synthesis and high-throughput screening of bound oligonucleotide and peptide sequences.

Fluorescent organosilica micro- and nanoparticles with controllable size

“…This swelling will cause the organosilica network porosity to increase, allowing the incorporation of large molecules such as organic dyes which covalently bind to the thiol functionalities of the organosilica. As shown in previous work, these organosilica particles retain their fluorescence when objected to various reagents involved in a multistep phosphoramidite oligonucleotide synthesis, whilst commercially available encoded polystyrene-divinylbenzene particles are unstable in most synthesis steps and lose most of their fluorescence [36]. These findings suggest that dye is covalently linked to the organosilica matrix, stopping the leaching of dye molecules during various oligonucleotide synthesis and washing steps.…”

“…Dithiocarbamate bonds are stable in acidic and neutral conditions, but easily cleaved in basic aqueous solution [35]. It is worth noting that for improved chemical stability maleimides can be used instead [36], which form a thioether bond that is not easily cleaved to yield free thiol [35]. Background fluorescence caused by free RBTIC dye was not detected, suggesting that the RBTIC-molecules are covalently coupled to the thiol groups and the washing steps were effective in removing excess dye.…”

“…Taking the size control and dye uptake studies as well as previous investigations [15,36] into consideration we propose a model for the particle formation process, as illustrated in Fig. 6.…”

Synthesis and application of streptavidin functionalized organosilica microparticles

“…27 And commercial polymer microparticles are typically optically encoded by physically entrapping, which are not unsuitable for organic solvent-based applications. 28,29 Therefore, home-made microparticles with uniform structure, good monodispersity of size, suitability for optical encoding, and availability in bulk, would be an ideal alternative. Organosilica microparticles achieves all of these objectives, which have been successfully synthesized and utilized in biomolecular screening, 28 DNA biosensors, 30 combinatorial synthesis, and microsphere-based flow cytometric immunoassays.…”

“…28,29 Therefore, home-made microparticles with uniform structure, good monodispersity of size, suitability for optical encoding, and availability in bulk, would be an ideal alternative. Organosilica microparticles achieves all of these objectives, which have been successfully synthesized and utilized in biomolecular screening, 28 DNA biosensors, 30 combinatorial synthesis, and microsphere-based flow cytometric immunoassays. 29 Glutaraldehyde has been one of the most potent and versatile tools for immobilizing of enzymes on a support.…”