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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

Experimentalsetups. (A) For the GAME configuration, the proteinis mixed with the PEG hydrogel and polymerized prior to the introductionof the capillary in the agarose gel. (B) For the 3L configuration,the protein–agarose or the protein–PEG (prepolymeric)solution is first loaded (first layer). For the latter, the protein–PEGsolution was subjected to UV treatment to induce the polymerization.This was followed by the gentle addition of the prepolymericsolution of PEG (20, 35, or 50% w/w concentrations), which was alsopolymerized by UV exposition (second layer or plug). The last stepin both experimental setups is the addition of the crystallizationsolution (third layer).
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fig1: Experimentalsetups. (A) For the GAME configuration, the proteinis mixed with the PEG hydrogel and polymerized prior to the introductionof the capillary in the agarose gel. (B) For the 3L configuration,the protein–agarose or the protein–PEG (prepolymeric)solution is first loaded (first layer). For the latter, the protein–PEGsolution was subjected to UV treatment to induce the polymerization.This was followed by the gentle addition of the prepolymericsolution of PEG (20, 35, or 50% w/w concentrations), which was alsopolymerized by UV exposition (second layer or plug). The last stepin both experimental setups is the addition of the crystallizationsolution (third layer).

Mentions: Counterdiffusionexperiments were set up using two configurations, the gel acupuncturemethod (GAME) with capillaries of different diameters (from 0.1 to0.8 mm) and the three-layer configuration (3L) using capillaries of2.5 mm inner diameter (Figure 1).


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)

Experimentalsetups. (A) For the GAME configuration, the proteinis mixed with the PEG hydrogel and polymerized prior to the introductionof the capillary in the agarose gel. (B) For the 3L configuration,the protein–agarose or the protein–PEG (prepolymeric)solution is first loaded (first layer). For the latter, the protein–PEGsolution was subjected to UV treatment to induce the polymerization.This was followed by the gentle addition of the prepolymericsolution of PEG (20, 35, or 50% w/w concentrations), which was alsopolymerized by UV exposition (second layer or plug). The last stepin both experimental setups is the addition of the crystallizationsolution (third layer).
© Copyright Policy
Related In: Results  -  Collection

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

fig1: Experimentalsetups. (A) For the GAME configuration, the proteinis mixed with the PEG hydrogel and polymerized prior to the introductionof the capillary in the agarose gel. (B) For the 3L configuration,the protein–agarose or the protein–PEG (prepolymeric)solution is first loaded (first layer). For the latter, the protein–PEGsolution was subjected to UV treatment to induce the polymerization.This was followed by the gentle addition of the prepolymericsolution of PEG (20, 35, or 50% w/w concentrations), which was alsopolymerized by UV exposition (second layer or plug). The last stepin both experimental setups is the addition of the crystallizationsolution (third layer).
Mentions: Counterdiffusionexperiments were set up using two configurations, the gel acupuncturemethod (GAME) with capillaries of different diameters (from 0.1 to0.8 mm) and the three-layer configuration (3L) using capillaries of2.5 mm inner diameter (Figure 1).

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