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Modified TMV particles as beneficial scaffolds to present sensor enzymes

Koch, C.; Wabbel, K.; Eber, F. J.; Krolla-Sidenstein, P. 1; Azucena, C. 1; Gliemann, H. 1; Eiben, S.; Geiger, F.; Wege, C.
1 Institut für Funktionelle Grenzflächen (IFG), Karlsruher Institut für Technologie (KIT)

Abstract:

Tobacco mosaic virus (TMV) is a robust nanotubular nucleoprotein scaffold increasingly employed for the high density presentation of functional molecules such as peptides, fluorescent dyes, and antibodies. We report on ist use as advantageous carrier for sensor enzymes. ATMV mutant with a cysteine residue exposed on every coat protein (CP) subunit (TMV$_{Cys}$) enabled the coupling of bifunctional maleimide-polyethylene glycol (PEG)-biotin linkers (TMV$_{Cys}$/Bio). Its surface was equipped with two streptavidin [SA]-conjugated enzymes: glucose oxidase ([SA]-GOx) and horseradish peroxidase ([SA]-HRP). At least 50% of the CPs were decorated with a linker molecule, and all thereof with active enzymes. Upon use as adapter scaffolds in conventional “high-binding” microtiter plates, TMV sticks allowed the immobilization of up to 45-fold higher catalytic activities than control samples with the same input of enzymes. Moreover, they increased storage stability and reusability in relation to enzymes applied directly to microtiter plate wells. The functionalized TMV adsorbed to solid supports showed a homogeneous distribution of the conjugated enzymes and structural integrity of the nanorods upon transmission electron and atomic force microscopy. ... mehr


Volltext §
DOI: 10.5445/IR/110103709
Originalveröffentlichung
DOI: 10.3389/fpls.2015.01137
Scopus
Zitationen: 78
Web of Science
Zitationen: 73
Dimensions
Zitationen: 82
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Funktionelle Grenzflächen (IFG)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2015
Sprache Englisch
Identifikator ISSN: 1664-462X
urn:nbn:de:swb:90-AAA1101037099
KITopen-ID: 110103709
HGF-Programm 47.02.06 (POF III, LK 01) Zellpopul.auf Biofunk.Oberflächen IFG
Erschienen in Frontiers in Plant Physiology
Verlag Frontiers Media SA
Band 6
Seiten 1137/1-17
Schlagwörter Tobacco mosaic virus(TMV), biotemplate, scaffold, glucose sensor, enzyme cascade system, enzyme shelf-life, reusability, surface immobilization
Nachgewiesen in Scopus
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Web of Science
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