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Biofunctionalization and self-interaction chromatography in PDMS microchannels

Paper ID Volume ID Publish Year Pages File Format Full-Text
3511 173 2012 9 PDF Available
Title
Biofunctionalization and self-interaction chromatography in PDMS microchannels
Abstract

In this paper we present an experimental protocol for protein immobilization on polydimethylsiloxane (PDMS) polymer surfaces and the subsequent application of a chromatographic PDMS microfluidic chip to measure protein–protein interactions. The PDMS surface modification steps are quantitatively and qualitatively experimentally analyzed using an array of techniques (water contact angle measurement, fluorescence spectroscopy and X-ray photoelectron spectroscopy). The protocol involves PDMS acidic surface activation using a potassium disulfite/potassium peroxidisulfate/acrylic acid mixture, followed by amination with 3-aminopropyl diethoxymethylsilane, followed by glutaraldehyde grafting and subsequent covalent protein binding. The applicability of such a miniaturized PDMS-based microfluidic system has been exemplified by measuring protein–protein interactions in a fast and accurate fashion for three model proteins, namely: hen egg white lysozyme, bovine ribonuclease-A and α-chymotrypsinogen. The protein interaction results align well with existing literature data using different materials and techniques. As the fabrication process for PDMS-based microstructures is relatively cheap, quick and requires limited lab expertise/access to specialized equipments, we consider that the implementation of such a flexible, easy to fabricate, PDMS-based microfluidic system for estimating protein interactions an important step toward quickly mapping protein phase behavior and measuring protein (self/cross) interactions in complex biological systems.

► We demonstrate a miniaturized flexible version of self-interaction chromatography by immobilizing proteins on polydimethylsiloxane (PDMS) microchannels. ► Fabrication process for PDMS-based microstructures is relatively cheap, quick and requires limited lab expertise. ► The resulting microchip is successfully applied and the protein interaction results align well with existing literature data. ► This approach will help to bring high throughput cheap microchip protein interaction screening within practical reach.

Keywords
Protein immobilization; Self-interaction chromatography; PDMS; Osmotic second virial coefficient; Microfluidic chip
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 67, 15 August 2012, Pages 111–119
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
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Any Questions? feel free to contact us