NDLI: Low-fouling SPR detection of lysozyme and its aggregates

Content Provider	Royal Society of Chemistry (RSC)
Author	Bucur, Bogdan Blaszykowski, Christophe Vasilescu, Alina Vezeanu, Alis Sheikh, Sonia David, Sorin Polonschii, Cristina Mihai, Iuliana Thompson, Michael Gáspár, Szilveszter
Copyright Year	2014
Abstract	Protein aggregates adsorb to material surfaces in a different manner than protein monomers and pose additional challenges for biosensor development with regard to non-specific adsorption (NSA). In this context, we describe herein the performance of a new antifouling thiol in a sensor coating resistant to NSA from lysozyme monomers and aggregates. Coatings were prepared as mixed self-assembling monolayers (SAMs) using a long polyethylene glycol carboxyl-terminated thiol (“PEG-COOH”) for the first time in conjunction with a shorter monoethylene glycol hydroxyl-terminated diluent thiol (“MEG-OH”). SAMs and their antifouling properties were characterized by a variety of surface analysis techniques. A key result was that the cleaning procedure drastically affects the antifouling properties of resulting MEG-OH based SAMs. Mixed PEG-COOH/MEG-OH SAMs formed on borohydride cleaned interfaces are able to reduce lysozyme NSA by >90% compared to bare gold; a remarkable performance also displayed for oligomers regardless of their stage of aggregation. Gratifyingly, subsequent SAM functionalisation with an anchoring layer of neutravidin for the preparation of a lysozyme sensor did not significantly alter the antifouling properties of the resulting assembly. The limit of detection for monomeric lysozyme by surface plasmon resonance was 0.3 μg mL−1 with a dynamic range of 3–50 μg mL−1 (R2 = 0.9993). The sensitivity of the technique for the aggregated lysozyme was almost two times higher than that for the protein monomer.
Starting Page	7646
Ending Page	7654
Page Count	9
File Format	HTM / HTML PDF
ISSN	17599660
Volume Number	6
Issue Number	19
Journal	Analytical Methods
DOI	10.1039/c4ay01237b
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Diluent Ethylene glycol Thiol Borohydride Dynamic range NeutrAvidin National Security Agency Surface plasmon Protein Adsorption Polyethylene glycol Biosensor Surface plasmon resonance Monomer NSA Lysozyme
Content Type	Text
Resource Type	Article
Subject	Analytical Chemistry Engineering Chemical Engineering

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