Covalent Biofunctionalization of Cellulose Acetate with Thermostable Chimeric Avidin

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 [(3)H]-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.

Multifunctional organic–inorganic hybrids based on cellulose acetate and 3-glycidoxypropyltrimethoxysilane

“…Additionally, pure CA films have also been grafted with 3-aminopropyltrimethoxysilane in order to introduce amino groups as a pathway to covalently immobilize proteins. In a recent work, Heikkinen et al [11] successfully evaluated the biotin-binding capacities of chimeric avidin, streptavidin, chicken avidin and rhizavidin immobilized on the surface of amino-functionalized CA films.…”

Aminosilane-Functionalized Cellulosic Polymer for Increased Carbon Dioxide Sorption

“…Cellulose acetate has been used for adsorption of a variety of chemical species, including phenyl compounds and polynuclear aromatic hydrocarbons (PAHs) in membrane separations, 9 N-methylcarbamates and lactose in ultrafiltration, 10 and N-isopropylacrylamide and N-n-butyl-acrylamide in membrane-based biochemistry immunoassay, 11 among many others. 12 Its CO 2 sorption properties and transport properties as CO 2 separation membranes have been previously investigated and fully characterized. 13 The results have shown great potential for this material to evolve into a promising CO 2 sorbent if a proper chemical treatment is applied in order to enhance its sorption properties.…”

Synthesis of Biopolymer‐Based Precursors for the Formation of Organic–Inorganic Hybrid Materials

“…Using CA, covalent bonding of 3‐aminopropyltrimethoxysilane and subsequent protein immobilization is known . The modification of CA with 3‐glycidoxypropyltrimethoxysilane could be used to incorporate luminescent Eu 3+ ‐complex or a highly branched alkoxysilane precursor was employed to determine the change in thermal‐ and dynamic‐mechanical properties of CA after crosslinking …”