Covalent Incorporation of Trehalose within Hydrogels for Enhanced Long-Term Functional Stability and Controlled Release of Biomacromolecules.

Covalent Incorporation of Trehalose within Hydrogels for Enhanced Long-Term Functional Stability and Controlled Release of Biomacromolecules.

Advanced healthcare materials (2015-06-20)

Timothy M O'Shea, Matthew J Webber, Alex A Aimetti, Robert Langer

ABSTRACT

Hydrogels with covalently incorporated trehalose are synthesized using thiol-ene Michael addition. Trehalose hydrogels afford prolonged stabilization and -controlled release of model enzymes in vitro and in vivo as well as preservation of protein stability under heat and -lyophilization stressors. Strong and -ordered hydrogen bonding interactions within covalently incorporated trehalose hydrogels represent a possible mechanism for protein stabilization.

MATERIALS

Product Number

Brand

Product Description

87748

Sigma-Aldrich

3,3′,5,5′-Tetramethylbenzidine, ≥98.0% (NT)

860336

Sigma-Aldrich

3,3′,5,5′-Tetramethylbenzidine, ≥99%

322415

Sigma-Aldrich

Methanol, anhydrous, 99.8%

517747

Sigma-Aldrich

Aluminum oxide, mesoporous, MSU-X (wormhole), average pore size 3.8 nm

W332615

Sigma-Aldrich

Acetone, natural, ≥97%

W332607

Sigma-Aldrich

Acetone, ≥99%, meets FCC analytical specifications

T2885

Sigma-Aldrich

3,3′,5,5′-Tetramethylbenzidine, ≥98% (TLC)

H9892

Sigma-Aldrich

Hydrochloric acid solution, 1.0 N, BioReagent, suitable for cell culture

T0167

Sigma-Aldrich

D-(+)-Trehalose dihydrate, from Saccharomyces cerevisiae, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99%

T9531

Sigma-Aldrich

D-(+)-Trehalose dihydrate, ≥99% (HPLC), from Saccharomyces cerevisiae

T5251

Sigma-Aldrich

D-(+)-Trehalose dihydrate, ≥98.5% (HPLC), from Saccharomyces cerevisiae

H1758

Sigma-Aldrich

Hydrochloric acid, 36.5-38.0%, BioReagent, for molecular biology

T9449

Sigma-Aldrich

D-(+)-Trehalose dihydrate, ≥99% (HPLC), from starch

T5525

Sigma-Aldrich

3,3′,5,5′-Tetramethylbenzidine, tablet, 1 mg substrate per tablet

705551

Sigma-Aldrich

Hydrogen chloride solution, 3 M in cyclopentyl methyl ether (CPME)

2104

Supelco

Hydrochloric acid solution, volumetric, 0.1 M HCl (0.1N), endotoxin free

L4777

Sigma-Aldrich

Lipase acrylic resin, ≥5,000 U/g, recombinant, expressed in Aspergillus niger

717975

Sigma-Aldrich

Methanol, NMR reference standard

84429

Sigma-Aldrich

Hydrochloric acid solution, ~6 M in H₂O, for amino acid analysis

W530574

Sigma-Aldrich

Hydrochloric acid solution, 32 wt. % in H₂O, FCC

270997

Sigma-Aldrich

Dichloromethane, anhydrous, ≥99.8%, contains 40-150 ppm amylene as stabilizer

202606

Sigma-Aldrich

Aluminum oxide, 99.997% trace metals basis

E34307

Sigma-Aldrich

Ethyl thioglycolate, 97%

537446

Sigma-Aldrich

Ethyl acetate, ReagentPlus^®, ≥99.8%

439223

Sigma-Aldrich

Dichloromethane, suitable for HPLC, ≥99.9%, contains 40-150 ppm amylene as stabilizer

429759

Sigma-Aldrich

Calcium chloride, AnhydroBeads^™, −10 mesh, ≥99.99% trace metals basis

41639

Sigma-Aldrich

Dimethyl sulfoxide, BioUltra, for molecular biology, ≥99.5% (GC)

W241407

Sigma-Aldrich

Ethyl acetate, ≥99%, FCC, FG

499609

Sigma-Aldrich

Calcium chloride, anhydrous, powder, 99.99% trace metals basis

449709

Sigma-Aldrich

Calcium chloride, AnhydroBeads^™, −10 mesh, ≥99.9% trace metals basis