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Covalent biofunctionalization of cellulose acetate with thermostable chimeric avidin

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Details

Original languageEnglish
Pages (from-to)2240-2245
Number of pages6
JournalACS Applied Materials and Interfaces
Volume3
Issue number7
DOIs
Publication statusPublished - 27 Jul 2011
Publication typeA1 Journal article-refereed

Abstract

A stable, bioactive cellulose acetate (CA) surface was developed by functionalizing the surface with highly thermostable avidin form. The CA films were first functionalized with a mixture of 3-aminopropyltrimethoxysilane and tetraethoxysilane to introduce free amino groups onto the surface of CA films. Free amino groups were functionalized with glutaraldehyde to obtain an activated surface for covalent biomolecule immobilization. A genetically engineered, high-affinity biotin-binding protein chimeric avidin, ChiAVD(I117Y), was used for biofunctionalization of the surface. The chimeric avidin protein has an increased stability in chemically harsh conditions and at high temperature when compared to wt (strept)avidin. The biological activity, i.e., biotin-binding capacity, of the immobilized protein was probed by [3H]-biotin. The activity of the chimeric avidin on functionalized CA films was fully retained over the three months' study period. The biotin-binding capacity of the immobilized chimeric avidin was compared to that of immobilized streptavidin, chicken avidin, and rhizavidin and significant differences between proteins were detected. The developed material offers a valuable platform for the development of inexpensive in vitro diagnostics and also supports biosensing applications that are required to operate under demanding conditions.

ASJC Scopus subject areas

Keywords

  • biomaterials, biotin, cellulose diacetate, chimeric avidin, glutaraldehyde, protein immobilization, sol-gel functionalization, thermostable protein