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

T5525

Sigma-Aldrich

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

T2885

Sigma-Aldrich

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

T0167

Sigma-Aldrich

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

2104

Supelco

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

T9531

Sigma-Aldrich

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

T5251

Sigma-Aldrich

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

87748

Sigma-Aldrich

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

L4777

Sigma-Aldrich

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

517747

Sigma-Aldrich

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

84429

Sigma-Aldrich

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

T9449

Sigma-Aldrich

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

W332607

Sigma-Aldrich

Acetone, ≥99%, meets FCC analytical specifications

W332615

Sigma-Aldrich

Acetone, natural, ≥97%

W530574

Sigma-Aldrich

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

705551

Sigma-Aldrich

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

860336

Sigma-Aldrich

3,3′,5,5′-Tetramethylbenzidine, ≥99%

717975

Sigma-Aldrich

Methanol, NMR reference standard

H1758

Sigma-Aldrich

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

H9892

Sigma-Aldrich

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

L092003

Sigma-Aldrich

Ethyl acetate

L092004

Sigma-Aldrich

Ethyl acetate

D4540

Sigma-Aldrich

Dimethyl sulfoxide, ≥99.5% (GC), suitable for plant cell culture

D1435

Sigma-Aldrich

Dimethyl sulfoxide, meets EP testing specifications, meets USP testing specifications

D8418

Sigma-Aldrich

Dimethyl sulfoxide, for molecular biology

D2650

Sigma-Aldrich

Dimethyl sulfoxide, Hybri-Max^™, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%

D2438

Sigma-Aldrich

Dimethyl sulfoxide, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma

C8106

Sigma-Aldrich

Calcium chloride

S7388

Sigma-Aldrich

N-Succinyl-Ala-Ala-Pro-Phe p-nitroanilide

C5670

Sigma-Aldrich

Calcium chloride, anhydrous, BioReagent, suitable for insect cell culture, suitable for plant cell culture, ≥96.0%

C3142

Sigma-Aldrich

α-Chymotrypsin from bovine pancreas, (TLCK treated to inactivate residual tryspin activity), Type VII, essentially salt-free, lyophilized powder, ≥40 units/mg protein