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The immobilization of proteins on biodegradable polymer fibers via click chemistry

Paper ID Volume ID Publish Year Pages File Format Full-Text
11262 727 2008 9 PDF Available
Title
The immobilization of proteins on biodegradable polymer fibers via click chemistry
Abstract

A facile and efficient method to immobilize bioactive proteins onto polymeric substrate was established. Testis-specific protease 50 (TSP50) was immobilized on ultrafine biodegradable polymer fibers, i.e., (1) to prepare a propargyl-containing polymer P(LA90–co–MPC10) by introducing propargyl group into a cyclic carbonate monomer (5-methyl-5-propargyloxycarbonyl-1,3-dioxan-2-one, MPC) and copolymerizing it with l-lactide; (2) to electrospin the functionalized polymer into ultrafine fibers; (3) to azidize the TSP50, and (4) to perform the click reaction between the propargyl groups on the fibers and the azido groups on the protein. The TSP50-immobilized fibers can resist non-specific protein adsorptions but preserve specific recognition and combination with anti-TSP50. ELISA tests were carried out by using HRP-goat-anti-mouse-IgG(H+L) as secondary antibody and o-phenylenediamine (OPDA)/H2O2 as substrate to detect the combination of immobilized TSP50 with anti-TSP50. The results showed that anti-TSP50 can be selectively adsorbed from its solution onto the TSP50-immobilized fibers in the presence of BSA of as high as 104 times concentration. TSP50 immobilized on the fiber and anti-TSP50 combined to the fiber were also quantitatively determined. Anti-TSP50 can be then eluted off from the fiber when pH changes. The eluted fiber can re-combine anti-TSP50 at an efficiency of 75% compared to the original TSP50-immobilized fiber. Therefore, the TSP50-immobilized fibers can be used in the detection, separation, and purification of anti-TSP50. The “click” method can lead to a universal strategy to protein immobilization.

Keywords
Click chemistry; TSP50; Protein immobilization; Biodegradable polymer; Electron spinning
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 29, Issue 8, March 2008, Pages 1118–1126
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
Online Support
Any Questions? feel free to contact us