A simple method for functionalization of cellulose membrane for covalent immobilization of biomolecules

被引:48
作者
Bora, U
Kannan, K
Nahar, P
机构
[1] CSIR, Inst Genom & Integrat Biol, Delhi 110007, India
[2] Guru Gobind Singh Indraprastha Univ, Sch Biotechnol, New Delhi 110006, India
关键词
cellulose membrane; photoactivation; covalent immobilization; enzyme; X-ray photoelectron spectroscopy;
D O I
10.1016/j.memsci.2004.10.028
中图分类号
TQ [化学工业];
学科分类号
0817 ;
摘要
Activated cellulose membrane was prepared by a simple photochemical reaction at 365 nm in 12 min using a photolinker, 1-fluoro-2-nitro-4-azidobenzene. XPS analysis of the activated cellulose membrane confirmed the presence of nitrogen and fluorine in the ratio of 2:1. Immobilization of a protein molecule onto the activated membrane occurred in 2 h at 37 degrees C. In contrast, no appreciable immobilization occurred onto the untreated surface. Disappearance of the fluorine peak in the XPS spectra of membrane having immobilized HRP confirmed covalent binding of the protein onto the activated membrane. Invertase was also immobilised onto the activated membrane and used in a flow through reactor system for conversion of sucrose to glucose and fructose. Immobilized invertase was found to be stable for at least 72 h of continuous run. The kinetic parameters of the enzyme reaction, Michaelis constant (K-m) and V-max, value of immobilized invertase was studied. The activated membrane when used in an ELISA procedure to detect immunoglobulins in human sera, showed around 2.6-fold higher sensitivity than the untreated membrane. The activated cellulose membrane has the potential for versatile applications such as in diagnostics, in flow reactor system for an enzyme-catalysed reaction and in membrane based affinity chromatography. (c) 2004 Elsevier B.V. All rights reserved.
引用
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页码:215 / 222
页数:8
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