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8 protocols using diethyl ether

1

Enzymatic Esterification of Polyols

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Lipase N435 derived from Candida antarctica (CAL-B) immobilized on acrylic resin (Sigma-Aldrich, St. Louis, MO, USA) was dried over P2O5 for 24 h prior to use. N,N-dimethylformamide (anhydrous, 99.8%), acetonitrile (anhydrous, 99.8%), tetrahydrofuran (anhydrous, 99.9%), xylitol (99%) and d-sorbitol (98%), stearoyl chloride (90%) and triethylamine were purchased from Sigma-Aldrich. Divinyl adipate (96%) was purchased from TCI GmbH (Eschborn, Germany). All chemicals were used without further purification. All other chemicals used in various stages like, diethyl ether, n-hexane, methanol and dichloromethane were purchased from Carl Roth (Karlsruhe, Germany).
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2

Synthesis of Photocurable Biomaterials

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Poly(ethylene glycol) (PEG) (Mn~10,000), PEG (Mn~400), Pentaerythritol ethoxylate (15/4 EO/OH) (Mn~797), methacrylic anhydride, acrylic acid, 4-dimethylaminopyridine (DMAP), PEGDA (Mn~575), isobornyl acrylate, diurethane dimethacrylate (DUDMA), diphenyl(2,4,6-trimethylbenzoyl) phosphine oxide (TPO), dibutyltin dilaurate, and pentaerythritol tetraacrylate (PETA) were purchased from Sigma-Aldrich (Taufkirchen, Germany). Tetrahydrofuran, diethyl ether, and 2-propanol were purchased from Carl Roth. Tinuvin-326 was kindly provided by BASF (Ludwigshafen, Germany).
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3

Synthesis and Characterization of Oxazoline Polymers

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Maleic anhydride (99%, Bernd Kraft), furan (99%, Acros), ethanolamine (99%, Acros), 4-toluenesulfonyl chloride (99%, Acros), 4-nitrobenzenesulfonyl chloride (97%, Sigma-Aldrich), pyridine (99%, Roth), tetramethylammonium hydroxide (25 wt% in methanol, Acros), bovine serum albumin (BSA; Sigma), toluene (Fisher), petroleum ether (Acros), ethanol (99.8%, Acros), acetone (Fisher), tetrahydrofuran (THF; VWR), diethyl ether (99.5%, Roth), and dichloromethane (DCM; VWR) were used as received. Acetonitrile (MeCN; Sigma-Aldrich) was dried in a solvent purification system (JC Meyer) before use as a polymerization solvent. 2-Ethyl-2-oxazoline (EtOx; Aldrich) was distilled over barium oxide and stored under argon.
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4

Reagent Preparation for Chemical Analysis

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Ethanol, methanol and n-hexane (all HPLC grade) were ordered from Th. Geyer (Renningen, Germany). Concentrated sulphuric acid (96-98%, for analysis p.a.) and diethyl ether (for synthesis, ≥ 99%) were from Carl Roth (Karlsruhe, Germany). Sodium chloride (≥ 98.9%), silver nitrate (≥ 99.5%, for analysis p.a.) and silica gel 60 were purchased from Sigma Aldrich (Steinheim, Germany). 2,2,4-Trimethylpentane (i-octane, for pesticide residue analysis) was ordered from Fluka Analytics (Seelze, Germany).
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5

Synthetic Methodology for Functionalized Biomaterials

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Dopamine hydrochloride,
sodium nitrite, sulfuric acid (98%), sodium azide, 2-bromoethylamine
hydrobromide, isobutyronitrile, ethanolamine, zinc acetate dihydrate,
anhydrous N,N-dimethylacetamide (DMA), copper bromine
(CuBr), sodium ascorbate, succinic anhydride, triethylamine (TEA),
4-(dimethylamino)pyridine (DMAP), N,N-diisopropylethylamine
(DIPEA), and anhydrous N,N-dimethylformamide (DMF)
were purchased from Merck KGaA (Darmstadt, Germany) and were used
without any purifications. Methyl-p-tosylate (MeTs)
and propargyl p-toluenesulfonate (PrTs) were purchased
from Merck KGaA (Darmstadt, Germany) and purified by distillation.
1-[(1-(Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylaminomorpholino)]uronium
hexafluorphosphate (COMU), potassium hydroxide (KOH), ethanol, diethyl
ether (Et2O), dichloromethane (DCM), hexane, and dry chloroform
were purchased from Carl Roth (Karlsruhe, Germany) and were used as
received. Dialysis tubes with different molecular weight cutoffs (3.5
kDa regenerated cellulose and 1000 kDa cellulose ester, Spectra/Por
Float-A-Lyzer) were purchased from Carl Roth.
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6

Analytical Grade Chemicals for LC-MS

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Methanol (MeOH) and acetonitrile (ACN) LC-MS grade and formic acid (FA) purity 99% were obtained from Carlo Erba Reagents (Chaussée du Vexin, France). Petroleum ether and diethyl ether analytical grade were purchased from Carl Roth GmbH (Karlsruhe, Germany), and deuterochloroform (CDCl 3 ) was obtained from Euriso-Top (Saint-Aubin, France). Potassium permanganate was obtained from Acros Organics (Geel, Belgium).
Ultrapure water was provided by a Purelab Classic (ELGA LabWater, High Wycombe, Bucks, UK).
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7

Analytical Reagents for Organic Synthesis

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Petroleum ether (40-60°C; 95%) was purchased from Acros Organics (Geel, Belgium). 2,3-Hexanedione (94%) and 2,4,5-trimethyl oxazole (97%) were obtained from Alfa Aesar (Karlsruhe, Germany).
Benzaldehyde (99%) and hydrogen peroxide (30%) were obtained from AppliChem (Darmstadt, Germany). Acetic acid (100%), chloroform-d (99.8 atom% D, with 0.03 vol% TMS, stabilized with Ag), diethyl ether (99.5%), iron(II) sulphate heptahydrate (99.5%), sodium carbonate decahydrate (98%), sodium sulphate (p.a.) and sulfuric acid (96%) were purchased from Carl Roth (Karlsruhe, Germany). Pentanal (97%) was obtained from Fisher Scientific (Darmstadt, Germany), Silica gel 60 was purchased from Macherey-Nagel (Düren, Germany) and 2,3-butanedione (97%) from Merck (Darmstadt, Germany). Acetoin (96%), 2,3-pentanedione (97%), 2,3,5,6-tetramethylpyrazine (98%) and 2,3,5-trimethylpyrazine (99%) were purchased from Sigma-Aldrich (Taufkirchen, Germany). 5-Ethyl-2,3-dimethylpyrazine (98%) was obtained from TCI (Eschborn, Germany). Butanal (97%) and propanal (97%) were purchased from Th. Geyer (Renningen, Germany) and ethyl acetate (98%) from VWR (Darmstadt, Germany). Helium (5.0) was obtained from Praxair (Düsseldorf, Germany) and nitrogen (5.0) from Air Liquide (Düsseldorf, Germany). All numbers given in parentheses represent the minimum purity.
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8

Polysaccharide-Based Multilayer Films

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Chitosan with deacetylation degree of 85% and average molecular weight, 𝑀 # ! , of 8.7 × 10 4 g/mol from Golden-Shell Biochemical (China), poly(ethylene imine) (PEI, 𝑀 # " = 2.5 × 10 4 g/mol) from Lupasol, BASF (Germany), and heparin sodium salt from Sigma-Aldrich (France) were used as polyelectrolytes. Methyl iodide and N-methyl-2-pyrrolidone, Dulbecco's modified eagle's medium (DMEM), penicillin 10,000 units/mL, streptomycin 10 mg/mL, MTT [(3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), and cellulose membrane (molecular weight cut off: 12,000 g/mol) used for dialysis, were purchased from Sigma-Aldrich (USA). Silicon wafers (orientation 100 and 0.5 mm in thickness) were obtained from WaferNet Inc. (EUA). Glass slides (diameter 15 mm, Menzel-Gläser) were used to build the LbL-films for cell culture assays. Other reactants, such as diethyl ether, isopropanol, sodium chloride, acetic acid, formaldehyde, and formic acid were purchased from Carl Roth (France) and used without further purification. Ultrapure water of Milli-Q quality (specific resistivity = 18.2 MΩ⋅cm at 25 °C) prepared from a water purification system (Millipore) was used for dialysis and for preparing aqueous solutions.
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