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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

Estimates of the diffusion coefficient of NaCl in cross-linkedPEG-based hydrogels of different compositions obtained by PS-MZI comparedwith the free water value (diamond). Error bars show 95% confidence.
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fig3: Estimates of the diffusion coefficient of NaCl in cross-linkedPEG-based hydrogels of different compositions obtained by PS-MZI comparedwith the free water value (diamond). Error bars show 95% confidence.

Mentions: Experimental results indicate that mesh size values varied fromapproximately 42 to 10 Å for 10 to 50% (w/w) composition, respectively.Moreover, the mesh size values for those hydrogels with 20–35%PEG composition did not show any significant statistical differences.In our case, the difference in dilution between these two compositionsmay be too small and measurements could fall within the margin oferror of the experimental protocol. However, the general trend observedis that when the monomer concentration in the prepolymeric solutionis reduced, there is an increase of mesh size. The effects of thesedifferences in the diffusion coefficient were evaluated by Mach–Zehnderinterferometer (Figure 3). A clear effecton the estimated diffusion coefficient of NaCl was observed for allpolymer compositions.


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)

Estimates of the diffusion coefficient of NaCl in cross-linkedPEG-based hydrogels of different compositions obtained by PS-MZI comparedwith the free water value (diamond). Error bars show 95% confidence.
© Copyright Policy
Related In: Results  -  Collection

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

fig3: Estimates of the diffusion coefficient of NaCl in cross-linkedPEG-based hydrogels of different compositions obtained by PS-MZI comparedwith the free water value (diamond). Error bars show 95% confidence.
Mentions: Experimental results indicate that mesh size values varied fromapproximately 42 to 10 Å for 10 to 50% (w/w) composition, respectively.Moreover, the mesh size values for those hydrogels with 20–35%PEG composition did not show any significant statistical differences.In our case, the difference in dilution between these two compositionsmay be too small and measurements could fall within the margin oferror of the experimental protocol. However, the general trend observedis that when the monomer concentration in the prepolymeric solutionis reduced, there is an increase of mesh size. The effects of thesedifferences in the diffusion coefficient were evaluated by Mach–Zehnderinterferometer (Figure 3). A clear effecton the estimated diffusion coefficient of NaCl was observed for allpolymer compositions.

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