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Use of Cross-Linked Poly(ethylene glycol)-Based Hydrogels for Protein Crystallization.

Gavira JA, Cera-Manjarres A, Ortiz K, Mendez J, Jimenez-Torres JA, Patiño-Lopez LD, Torres-Lugo M - Cryst Growth Des (2014)

Bottom Line: PEG hydrogels also induced the nucleation of lysozyme crystals to a higher extent than agarose.As an example, insulin crystals were grown in 10% (w/w) PEG hydrogel.The resulting crystals were of an approximate size of 500 μm.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Estudios Crystalográficos, IACT (CSIC-UGR). Avda. las Palmeras 4, E18100 Armilla, Granada, Spain.

ABSTRACT
Poly(ethylene glycol) (PEG) hydrogels are highly biocompatible materials extensively used for biomedical and pharmaceutical applications, controlled drug release, and tissue engineering. In this work, PEG cross-linked hydrogels, synthesized under various conditions, were used to grow lysozyme crystals by the counterdiffusion technique. Crystallization experiments were conducted using a three-layer arrangement. Results demonstrated that PEG fibers were incorporated within lysozyme crystals controlling the final crystal shape. PEG hydrogels also induced the nucleation of lysozyme crystals to a higher extent than agarose. PEG hydrogels can also be used at higher concentrations (20-50% w/w) as a separation chamber (plug) in counterdiffusion experiments. In this case, PEG hydrogels control the diffusion of the crystallization agent and therefore may be used to tailor the supersaturation to fine-tune crystal size. As an example, insulin crystals were grown in 10% (w/w) PEG hydrogel. The resulting crystals were of an approximate size of 500 μm.

No MeSH data available.


Related in: MedlinePlus

Lysozyme crystals grownat constant temperature in PEG hydrogelswith different polymer compositions: 5% (w/w) (top left), 7% (w/w)(top right), 9% (w/w) (bottom left), and 10% (w/w) (bottom right).
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fig5: Lysozyme crystals grownat constant temperature in PEG hydrogelswith different polymer compositions: 5% (w/w) (top left), 7% (w/w)(top right), 9% (w/w) (bottom left), and 10% (w/w) (bottom right).

Mentions: Lysozymecrystals were successfully grown in 0.8 mm capillaries in the presenceof PEG hydrogels at different polymer concentrations (i.e., degreesof monomer dilutions prior to polymerization). A monomer solutioncontaining a 1:1 molar ratio of monomer and cross-linker was furtherdiluted with the appropriate protein solution to obtain a monomerconcentration ranging from 5 to 10% (w/w). Crystals grown at 5 and7% hydrogel concentration showed the typical shape of lysozyme crystalscomposed of prismatic (110) and pyramidal (001) faces, while at 9%(w/w), crystals showed a rounded shape that became almost sphericalat 10% (w/w) (Figure 5).


Use of Cross-Linked Poly(ethylene glycol)-Based Hydrogels for Protein Crystallization.

Gavira JA, Cera-Manjarres A, Ortiz K, Mendez J, Jimenez-Torres JA, Patiño-Lopez LD, Torres-Lugo M - Cryst Growth Des (2014)

Lysozyme crystals grownat constant temperature in PEG hydrogelswith different polymer compositions: 5% (w/w) (top left), 7% (w/w)(top right), 9% (w/w) (bottom left), and 10% (w/w) (bottom right).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215911&req=5

fig5: Lysozyme crystals grownat constant temperature in PEG hydrogelswith different polymer compositions: 5% (w/w) (top left), 7% (w/w)(top right), 9% (w/w) (bottom left), and 10% (w/w) (bottom right).
Mentions: Lysozymecrystals were successfully grown in 0.8 mm capillaries in the presenceof PEG hydrogels at different polymer concentrations (i.e., degreesof monomer dilutions prior to polymerization). A monomer solutioncontaining a 1:1 molar ratio of monomer and cross-linker was furtherdiluted with the appropriate protein solution to obtain a monomerconcentration ranging from 5 to 10% (w/w). Crystals grown at 5 and7% hydrogel concentration showed the typical shape of lysozyme crystalscomposed of prismatic (110) and pyramidal (001) faces, while at 9%(w/w), crystals showed a rounded shape that became almost sphericalat 10% (w/w) (Figure 5).

Bottom Line: PEG hydrogels also induced the nucleation of lysozyme crystals to a higher extent than agarose.As an example, insulin crystals were grown in 10% (w/w) PEG hydrogel.The resulting crystals were of an approximate size of 500 μm.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Estudios Crystalográficos, IACT (CSIC-UGR). Avda. las Palmeras 4, E18100 Armilla, Granada, Spain.

ABSTRACT
Poly(ethylene glycol) (PEG) hydrogels are highly biocompatible materials extensively used for biomedical and pharmaceutical applications, controlled drug release, and tissue engineering. In this work, PEG cross-linked hydrogels, synthesized under various conditions, were used to grow lysozyme crystals by the counterdiffusion technique. Crystallization experiments were conducted using a three-layer arrangement. Results demonstrated that PEG fibers were incorporated within lysozyme crystals controlling the final crystal shape. PEG hydrogels also induced the nucleation of lysozyme crystals to a higher extent than agarose. PEG hydrogels can also be used at higher concentrations (20-50% w/w) as a separation chamber (plug) in counterdiffusion experiments. In this case, PEG hydrogels control the diffusion of the crystallization agent and therefore may be used to tailor the supersaturation to fine-tune crystal size. As an example, insulin crystals were grown in 10% (w/w) PEG hydrogel. The resulting crystals were of an approximate size of 500 μm.

No MeSH data available.


Related in: MedlinePlus