title: Versatile bio-ink for covalent immobilization of chimeric avidin on sol-gel substrates.
abstract: A bio-ink for covalent deposition of thermostable, high affinity biotin-binding chimeric avidin onto sol-gel substrates was developed. The bio-ink was prepared from heterobifunctional crosslinker 6-maleimidohexanoic acid N-hydroxysuccinimide which was first reacted either with 3-aminopropyltriethoxysilane or 3-aminopropyldimethylethoxysilane to form silane linkers 6-maleimide-N-(3-(triethoxysilyl)propyl)hexanamide or -(ethoxydimethylsilyl)propyl)-hexanamide. C-terminal cysteine genetically engineered to chimeric avidin was reacted with the maleimide group of silane linker in methanol/PBS solution to form a suspension, which was printed on sol-gel modified PMMA film. Different concentrations of chimeric avidin and ratios between silane linkers were tested to find the best properties for the bio-ink to enable gravure or inkjet printing. Bio-ink prepared from 3-aminopropyltriethoxysilane was found to provide the highest amount of active immobilized chimeric avidin. The developed bio-ink was shown to be valuable for automated fabrication of avidin-functionalized polymer films.
authors.affiliation: Department of Chemistry, University of Oulu, Oulu, Finland. jarkko.j.heikkinen@oulu.fi
authors.forename: Jarkko J
authors.initials: JJ
authors.lastname: Heikkinen
authors.forename: Liisa
authors.initials: L
authors.lastname: Kivimäki
authors.forename: Juha A E
authors.initials: JA
authors.lastname: Määttä
authors.forename: Inka
authors.initials: I
authors.lastname: Mäkelä
authors.forename: Leena
authors.initials: L
authors.lastname: Hakalahti
authors.forename: Kristiina
authors.initials: K
authors.lastname: Takkinen
authors.forename: Markku S
authors.initials: MS
authors.lastname: Kulomaa
authors.forename: Vesa P
authors.initials: VP
authors.lastname: Hytönen
authors.forename: Osmo E O
authors.initials: OE
authors.lastname: Hormi
mesh_terms: D001360:Avidin / Q000235:genetics / Q000378:metabolism*; D001672:Biocompatible Materials / Q000138:chemical synthesis* / Q000378:metabolism; D001710:Biotin / Q000737:chemistry / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents / Q000737:chemistry; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005782:Gels / Q000737:chemistry* / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007281:Ink; D044367:Phase Transition; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000737:chemistry / Q000378:metabolism; D011437:Propylamines; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012821:Silanes / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry; D013499:Surface Properties
chemical_list: D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D005782:Gels; D055767:Immobilized Proteins; D011108:Polymers; D011437:Propylamines; D011994:Recombinant Proteins; D012821:Silanes; D013388:Succinimides; D001360:Avidin; D001710:Biotin; D003545:Cysteine; C477625:amino-propyl-triethoxysilane; C001426:N-hydroxysuccinimide