Ethanolamine hydrochloride

Peptide sequences employed in nanoMIP development
(Ontores Biotechnology, Zhejiang, China), monomers for nanoMIP synthesis
(Sigma, Gillingham, UK), sodium hydroxide (Sigma), ammonium persulfate
(Sigma), mercaptoundecanoic acid (Sigma), ethanolamine hydrochloride
(Sigma), bovine serum albumin (BSA, Sigma), Pluronic solution (Sigma),
4-ABA (Fisher Scientific, Loughborough, UK), EDC (Fisher Scientific),
NHS (Fisher Scientific), ferricyanide (Sigma), ferrocyanide (Sigma),
potassium chloride (KCl, Sigma), hydrogen peroxide (30%, Sigma), ammonia
solution (25%, VWR International, Leicestershire, UK), PBS tablets
(Sigma), the alpha variant of the SARS-CoV-2 spike protein (The Native
Antigen Company, Kidlington, UK), the delta variant of the spike protein
(Abbexa, Cambridge, UK), HSA (Sigma), and IL-6 (Bio-Rad, Watford,
UK) were used as received. The SARS-CoV-2 RBD and ORF8 were provided
by the Medical Research Council Protein Phosphorylation and Ubiquitylation
Unit (Dundee, UK). PBS solutions were prepared with deionized (DI)
water, and AFM experiments were performed using Milli-Q water (both
with a resistivity of ≥18.2 MΩ cm).

Cyanogen bromide activated Sepharose 4B (CNBr) and butyl Sepharose CL-8B were purchased from General Electric (Upsala, Sweden). Agarose 10BCL (50–150 μm) was purchased from Agarose Bead Technologies (Madrid, Spain). Fully activated glyoxyl–agarose 10BCL (150 μmol aldehyde groups/g) was prepared as previously described [30 (link)]. Thermomyces lanuginosus lipase (TLL), Candida antarctica lipase sp. 99–125 (CAL), 1,2-ethylenediamine (EDA), Ethanolamine hydrochloride, sodium metaperyodate, 1-ethyl-3-(dimethylaminopropyl) carbodiimide (EDC), Triton X-100, dithiotreitol (DTT), anthranilic acid (AA), methyl anthranilate (MA), aniline (AN), ethanol, p-nitrophenyl butyrate (p-NPB), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA) and salts for buffering solutions were purchased from Sigma Chem. Co. (St. Louis, MO, USA). The sardine oil was a gift from BTSA, Biotecnologías Aplicadas, S.L. (Madrid, Spain); the Novozyme^® 435 was a gift from Novozymes (Bagsværd, Denmark). Geobacillus thermocatenulathus lipase 2 (BTL2) expressed in E. coli was produced, purified, and aminated in solid phase as previously described [22 (link)]. Other reagents and solvents were of analytical or HPLC grade. Novozyme^® 435 and Lewatit^® VP OC 1600 were kindly donated by Novozymes A/S and Lanxess^®, respectively.

The laccase enzymes from Rhus vernicifera and salts employed in buffer solutions were purchased from Sigma-Aldrich (St. Louis, MO, USA). The chemical compounds 3-Phenoxybenzaldehyde, 16-mercaptohexadecanoic acid (MHDA), 11-mercaptoundecanol (MUD), ethanolamine hydrochloride, N-hydroxysuccinimide (NHS), and 1-thyl-3-(3-dimethylamino-ropyl) carbodiimide hydrochloride (EDC) were supplied from Sigma-Aldrich (St. Louis, MO, USA). The stock solutions of 3-Phenoxybenzaldehyde (100 mg·mL⁻¹) were prepared in ethanol:water (90:10, % V/V), and from them, dilutions were prepared in phosphate buffer solution (PBS), pH 7.3 (0.1 M). Chrome and gold pellets (99.999%) were purchased from Kurt J. Lesker Co. (Clairton, PA, USA) and employed in the thin-film process’s evaporation.

Ethanolamine hydrochloride, tetrahydrofuran (THF), dichloromethane, dimethylsulfoxide (DMSO), sodium cyanoborohydride, acetone, ethanol, bovine serum albumin (BSA), o-phenylenediamine (OPD), laminarin from Laminaria digitate, α-mannan from Saccharomyces cerevisiae (M75014), d-glucose, d-mannose, N-acetyl-d-glucosamine were purchased form Sigma-Aldrich and used without further purification. Pustulan (β-1,6-glucan) and β-1,6-gentiobiose were obtained from Calbiochem and Carbosynth, respectively. β-1,3-laminaribiose and penta-N-acetylchitopentaose were provided by Megazyme. PBS buffer (10 mM Na₂HPO₄, 138 mM NaCl, 2.7 mM KCl, pH 7.4) was purchased in Euromedex. Epoxide-activated glass slides were from Schott Nexterion (Nexterion® Slide E). The MLCT AFM tips were obtained from Bruker. Wheat Germ Agglutinin (WGA) and concanavalin A (ConA) from Sigma-Aldrich. Anti-β-1,3-glucan, a mouse monoclonal IgG was provided by Biosupplies (http://www.biosupplies.com.au/, Australia). The anti-β-1,6-glucan antibody, a rabbit polyclonal antibody IgG, was kindly provided by Prof. Frans Klis (Swammerdam University of Amsterdam). A goat-anti-rabbit and a rabbit anti-mouse IgG-peroxidase conjugates were purchased from ThermoFisher Scientific.