Bar-coded hydrogel microparticles for protein detection: synthesis, assay and scanning
| Autor: | Stephen C. Chapin, Rathi L. Srinivas, Patrick S. Doyle, David C. Appleyard |
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| Rok vydání: | 2011 |
| Předmět: |
Range (particle radiation)
Electronic Data Processing Microscopy Materials science Chromatography Microfluidics Proteins Hydrogels Serum Albumin Bovine Molecular biology General Biochemistry Genetics and Molecular Biology Microspheres Suspension (chemistry) chemistry.chemical_compound chemistry Particle Animals Wafer Cattle Microparticle Ethylene glycol Lithography Antibodies Immobilized |
| Zdroj: | Nature protocols. 6(11) |
| ISSN: | 1750-2799 |
| Popis: | This protocol describes the core methodology for the fabrication of bar-coded hydrogel microparticles, the capture and labeling of protein targets and the rapid microfluidic scanning of particles for multiplexed detection. Multifunctional hydrogel particles made from poly(ethylene glycol) serve as a sensitive, nonfouling and bio-inert suspension array for the multiplexed measurement of proteins. Each particle type bears a distinctive graphical code consisting of unpolymerized holes in the wafer structure of the microparticle; this code serves to identify the antibody probe covalently incorporated throughout a separate probe region of the particle. The protocol for protein detection can be separated into three steps: (i) synthesis of particles via microfluidic flow lithography at a rate of 16,000 particles per hour; (ii) a 3-4-h assay in which protein targets are captured and labeled within particles using an antibody sandwich technique; and (iii) a flow scanning procedure to detect bar codes and quantify corresponding targets at rates of 25 particles per s. By using the techniques described, single- or multiple-probe particles can be reproducibly synthesized and used in customizable multiplexed panels to measure protein targets over a three-log range and at concentrations as low as 1 pg ml(-1). |
| Databáze: | OpenAIRE |
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