Rehydratable gel for rapid loading of nanoliter solution and its application in protein crystallization

Li, Yuefang; Guo, Dameng; Zheng, Bo

doi:10.1039/c2ra20511d

Cited by 8 publications

(11 citation statements)

References 34 publications

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“…Also, the conventional ways to delivery proteins and precipitants inside the gel network, to create supersaturation, and crystals recovery might include several and uneasy procedures. [ 14 ] On these basis, the fundamental idea of this study is to improve current approaches to protein crystallization by developing functional structured materials (composite or hydrogel membranes) useful to combine the general advantages of crystallization in gel (reproducibility, size increase, and mechanical stability of crystals) with those of membrane-assisted undesired in order to avoid high-resistance to mass transport in solvent evaporation membrane crystallization. [ 15 ] Accordingly, hydrogel membranes were fabricated in three main steps ( Figure 1 a): 1) conditioning of PP supports to swell the polymer and to enhance its chemical affi nity with the aqueous pre-polymerization solution; 2) casting of a homogeneous layer of the pre-polymerization solution above the conditioned membrane, preventing solution intrusion inside the pores; 3) rapid polymerization of the solution under the UV lamp with the formation of the gel layer ( Figure 2 a,b, upper part).…”

Section: Introductionmentioning

confidence: 99%

Tailored Hydrogel Membranes for Efficient Protein Crystallization

Profio

Polino

Nicoletta

et al. 2013

Adv Funct Materials

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Crystallization still represents the bottleneck in the process of protein structure determination at high resolution, despite high-throughput structural genomics programs requiring optimized crystallization strategies regarding crystal quality, time, success rate, reproducibility, and used protein amount. On the other hand, the development of suitable materials for controlled heterogeneous nucleation might facilitate biomacromolecular crystallization in a variety of experimental conditions which are not conventionally fruitful. Here, the possibility to fabricate hydrogel membranes displaying controlled chemical composition and nanostructure and to use them as heterogeneous supports for biomacromolecular crystallization is demonstrated. Diverse gel morphologies are obtained by controlling phase separation kinetics during gel layer formation on membrane support. These composite materials are found to increase the effi ciency of the crystallization process so that crystals with enhanced diffraction properties are produced at lower protein concentration than conventional technique, thus affording the possibility to improve current approaches to protein crystallization and to be adapted to specifi c targets.

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Section: Introductionmentioning

confidence: 99%

Tailored Hydrogel Membranes for Efficient Protein Crystallization

Profio

Polino

Nicoletta

et al. 2013

Adv Funct Materials

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“…The PURE system proteinaceous factors, ribosome and plasmid templates were loaded into the PA hydrogel particles when the predried PA hydrogel particles were rehydrated in the PURE system solution. 45 The enzyme mixture of the PURE system includes 20 tRNA synthetases, 3 initiation factors (IF1, IF2, and IF3), 3 elongation factors (EF-G, EF-Tu, and EF-Ts), 3 release factors (RF1, RF2, and RF3), ribosome recycling factor, myokinase, creatine kinase, nucleoside-diphosphate kinase, peptidylprolyl-isomerase, T7 RNA polymerase, and 70S ribosome. The proteinaceous factors of the PURE system were all labelled with the His-tag and immobilized through the interaction of the His-tag and the Ni 2+ –NTA complex.…”

Section: Resultsmentioning

confidence: 99%

Long-lived protein expression in hydrogel particles: towards artificial cells

Zhou

Cui

et al. 2018

Chem. Sci.

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“…The microdevice was validated by performing the Michaelis-Menten kinetics analysis of alkaline phosphatase. The microdevice is simple, inexpensive and easy to operate, and is suitable for serial assays and screening 10,39 in bioanalysis.…”

Section: Discussionmentioning

confidence: 99%

A pneumatic valve controlled microdevice for bioanalysis

Zhou

Zheng

2013

Biomicrofluidics

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This paper describes a pneumatic valve controlled microdevice for performing mixing and reaction. This microdevice combined the degassed polydimethylsiloxane (PDMS) pumping method with a syringe-actuated valve system to control the dispensing and mixing of nanoliter solutions. The syringe was used to manually generate vacuum and to open the valves. Upon the opening of the valve, the microchamber was filled with the solution, which was driven by the external atmosphere through the degassed PDMS microchannel. With this microdevice, the enzymatic kinetics of alkaline phosphatase converting the fluorescein diphosphate was studied, and the Michaelis-Menten kinetics was analyzed. The microdevice has the advantages of simplicity and low cost in fabrication and operation.

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Rehydratable gel for rapid loading of nanoliter solution and its application in protein crystallization

Cited by 8 publications

References 34 publications

Tailored Hydrogel Membranes for Efficient Protein Crystallization

Tailored Hydrogel Membranes for Efficient Protein Crystallization

Long-lived protein expression in hydrogel particles: towards artificial cells

A pneumatic valve controlled microdevice for bioanalysis

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