Generic Method for Attaching Biomolecules via Avidin–Biotin Complexes Immobilized on Films of Regenerated and Nanofibrillar Cellulose

Abstract

We investigated the adsorption and chemical conjugation of avidin and its deglycosylated form, neutravidin, on films of regenerated and nanofibrillar cellulose. The dynamics and extent of biomolecular attachment were monitored in situ by quartz crystal microbalance microgravimetry and ex situ via surface analyses with atomic force microscopy and X-ray photoelectron spectroscopy. The installation of carboxyl groups on cellulose after modification with carboxymethylated cellulose (CMC) or TEMPO-oxidation significantly increases physisorption of avidins, which can be then covalently conjugated by using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride/N-hydroxysuccinimide (EDS/NHS) coupling chemistries. The developed cellulose–avidin biointerfaces are able to scavenge biotinylated molecules from solution as demonstrated by successful surface complexation of biotinylated bovine serum albumin (Biotin-BSA) and antihuman immunoglobulin G (Biotin-anti-hIgG). Finally, we show that cellulose substrates carrying immobilized anti-hIgG are effective in detecting human immunoglobulin G (hIgG) from fluid matrices.

Homogeneous cellulose thin films by regeneration of cellulose xanthate: properties and characterization

“…Therefore, they have been proposed as supports for biosensors, as matrix for catalysis as well as material for optoelectronic devices such as transistors and solar cells (Blomstedt et al 2007;Filpponen et al 2012;Niegelhell et al 2016;Orelma et al 2011Orelma et al , 2012Reishofer et al 2017;Taajamaa et al 2013;Wolfberger et al 2015). Further, these films have been employed as models to investigate and to better understand interactions of water with cellulose, particularly in the context of cell wall structure and their relation to industrially applied drying processes (Ehmann et al 2015;Niinivaara et al 2015Niinivaara et al , 2016.…”

Tailor-made functional surfaces based on cellulose-derived materials

“…Physical adsorptions are usually weak and reversible. However, the adsorption of carboxymethyl cellulose (CMC) onto cellulose surfaces is generally specific and irreversible, (Kargl et al 2012), and thus CMC is widely used as anchoring layers on cellulose surfaces for further modifications Mohan et al 2014;Orelma et al 2011;Orelma et al 2012b;Orelma et al 2012c). Covalent modification can be realized via Bgraft onto^and Bgraft from^approaches.…”

“…In the graft onto approach, bioactive molecules or polymers are coupled to the hydroxyl, carboxylic acid, or other functional groups on cellulose surfaces via covalent bond formation. Commonly used graft onto chemistries include azide-alkyne cycloaddition (Krouit et al 2008;Pahimanolis et al 2011), thiol-ene coupling (Tingaut et al 2011;Zhao et al 2010), c o u p l i n g r e a c t i o n s a c t i v a t e d b y 1 -e t h y l -3 -( 3 -dimethylaminopropyl)carbodiimid (EDC) and Nhydroxysuccinimide (NHS) (Arola et al 2012;Orelma et al 2012a;Orelma et al 2012b), oxime ligation (Uth et al 2014), bifunctional reagent-mediated crosslinking (e.g., diisothiocyanate (Araújo et al 2012) and divinyl sulfone (Sannino et al 2003;Yu et al 2012)), Diels-Alder cycloaddition (Tischer et al 2012), photo-induced crosslinking Dietrich et al 2012), and so on. In the graft from approach, initiators are immobilized onto cellulose Scheme 1 Overview of the preparation of functional surfaces using cellulose-based materials surfaces and monomers are polymerized directly from the cellulose surfaces via surface-initiated polymerizations (i.e., atom transfer radical polymerization (ATRP)) Nystrom et al 2006;Qiu et al 2013), reversible addition-fragmentation transfer polymerization (RAFT) , ring-opening polymerization (ROP) (Boujemaoui et al 2012;Goffin et al 2011), and ringopening metathesis polymerization (ROMP) .…”

“…In the graft from approach, initiators are immobilized onto cellulose Scheme 1 Overview of the preparation of functional surfaces using cellulose-based materials surfaces and monomers are polymerized directly from the cellulose surfaces via surface-initiated polymerizations (i.e., atom transfer radical polymerization (ATRP)) Nystrom et al 2006;Qiu et al 2013), reversible addition-fragmentation transfer polymerization (RAFT) , ring-opening polymerization (ROP) (Boujemaoui et al 2012;Goffin et al 2011), and ringopening metathesis polymerization (ROMP) . For a biochemical coupling, bioactive molecules or nanoparticles are immobilized onto cellulose surfaces (or modified cellulose surfaces) via carbohydrate-binding module containing bifunctional fusion proteins (Pangu et al 2007;Ye et al 2009), ligand-receptor pairing (e.g., avidin-biotin interaction) (Orelma et al 2012b), and enzyme-mediated ligation (Uth et al 2014). …”

“…Thus, CMC could be utilized as a linker molecule for bioactive molecule immobilization onto cellulose surfaces. Orelma et al, for instance, reported the immobilization of antibodies onto regenerated and nanofibrillar cellulose surfaces using CMC as a linker molecule (Orelma 2012;Orelma et al 2012b;Orelma et al 2012c). Anti-hemoglobin was covalently coupled to CMCmodified cellulose surfaces via EDC/NHS chemistry.…”