Refolding of proteins must be performed under very dilute conditions to overcome the competing aggregation reaction, which has a high reaction order. Refolding on a chromatography column partially prevents formation of the intermediate form prone to aggregation. A chromatographic refolding procedure was developed using an autoprotease fusion protein with the mutant EDDIE from the Npro autoprotease of pestivirus. Upon refolding, self-cleavage generates a target peptide with an authentic N-terminus. The refolding process was developed using the basic 1.8-kDa peptide sSNEVi-C fused to the autoprotease EDDIE or the acidic peptide pep6His, applying cation and anion exchange chromatography, respectively. Dissolved inclusion bodies were loaded on cation exchange chromatographic resins (Capto S, POROS HS, Fractogel EMD SO3−, UNOsphere S, SP Sepharose FF, CM Sepharose FF, S Ceramic HyperD F, Toyopearl SP-650, and Toyopearl MegaCap II SP-550EC). A conditioning step was introduced in order to reduce the urea concentration prior to the refolding step. Refolding was initiated by applying an elution buffer containing a high concentration of Tris–HCl plus common refolding additives. The actual refolding process occurred concurrently with the elution step and was completed in the collected fraction. With Capto S, POROS HS, and Fractogel SO3−, refolding could be performed at column loadings of 50 mg fusion protein/ml gel, resulting in a final eluate concentration of around 10–15 mg/ml, with refolding and cleavage step yields of around 75%. The overall yield of recovered peptide reached 50%. Similar yields were obtained using the anion exchange system and the pep6His fusion peptide. This chromatographic refolding process allows processing of fusion peptides at a concentration range 10- to 100-fold higher than that observed for common refolding systems. 相似文献
on the hyperbolic spaces \({\mathbb{H}^d}\), d ≥ 2, for exponents \({\sigma \in (0, 2/(d-2))}\). The main unexpected conclusion is scattering to linear solutions in the case of small exponents σ; for comparison, on Euclidean spaces scattering in H1 is not known for any exponent \({\sigma \in (1/d, 2/d]}\) and is known to fail for \({\sigma \in (0, 1/d]}\). Our main ingredients are certain noneuclidean global in time Strichartz estimates and noneuclidean Morawetz inequalities.
Single-walled carbon nanotubes (SWNTs) are 1D nanostructures with distinct physical and chemical properties that have shown great promise for applications in many fields, including biomedicine. Since for biomedical application the water solubility is crucial and SWNTs have low solubility, various methods (including polymer and biopolymer wrapping, chemical modifications) have been developed to solubilize and disperse them in water. Due to their unique optical properties such as photoluminescence in the NIR and strong resonant Raman signatures, they can be used as nanoprobes in biomedical imaging and phototherapies. Furthermore, decoration of SWNTs with noble metal nanoparticles will induce an excellent surface-enhanced Raman scattering (SERS) effect of the nanoparticles-SWNTs composites, with applications in cell imaging. Herein, we present a new and facile strategy for the DNA-assisted decoration of SWNTs with gold nanoparticles (AuNPs) and their application in SERS imaging. By ultrasonication at room temperature of SWNTs with AuNPs functionalized with synthetic DNA, SWNT-AuNPs nanocomposites with enhanced Raman signal were obtained. Among the important advantages of the proposed method are the presence of the free DNA overhangs around the SWNT-AuNPs suitable for post-synthetic modification of nanocomposite through hybridization of complementary DNA strands containing molecules of interest attached by well-developed bio-conjugation chemistry.
Applications requiring pristine graphene derived from graphite demand a solution stabilization method that utilizes an easily removable media. Using a combination of molecular dynamics simulations and experimental techniques, we investigate the solublization/suspension of pristine graphene sheets by an equimolar mixture of benzene and hexafluorobenzene (C(6)H(6)/C(6)F(6)) that is known to form an ordered structure solidifying at 23.7 °C. Our simulations show that the graphene surface templates the self-assembly of the mixture into periodic layers extending up to 30 ? from both sides of the graphene sheet. The solvent structuring is driven by quadrupolar interactions and consists of stacks of alternating C(6)H(6)/C(6)F(6) molecules rising from the surface of the graphene. These stacks result in density oscillations with a period of about 3.4 ?. The high affinity of the 1:1 C(6)H(6)/C(6)F(6) mixture with graphene is consistent with observed hysteresis in Wilhelmy plate measurements using highly ordered pyrolytic graphite (HOPG). AFM, SEM, and TEM techniques verify the state of the suspended material after sonication. As an example of the utility of this mixture, graphene suspensions are freeze-dried at room temperature to produce a sponge-like morphology that reflects the structure of the graphene sheets in solution. 相似文献
We prove a bifurcation result for a perturbed sublinear elliptic problem defined on a compact Riemannian manifold without boundary. This result is then applied to a singular elliptic problem on even-dimensional Euclidean spaces. 相似文献
Perpendicular structured nanowire arrays interaction field distributions (IFDs), as revealed from first-order reversal curves (FORC) diagrams, are related to the presence of the demagnetizing field in the system. Despite the similarity between the geometric properties of bit patterned media and mentioned nanowire arrays, FORC diagrams of these two types of systems are not similar essentially due to the different number of magnetic entities influencing the switch of an individual element. We show that one Preisach–Krasnosel'skii–Pokrovskii (PKP) symmetrical hysteron can be representative of an ideal infinite nanowire array when the field is applied along the wires. Starting from this observation, we present a very simple model based on PKP symmetrical hysterons that can be applied to real finite ferromagnetic nanowire arrays, and is able to describe a wide class of experimentally observed FORC distributions, revealing features due to size effects. We also present IFDs modeled for different geometric characteristics such as array size, interwire distance, and nanowire dimensions, and an identification procedure for the proposed model. 相似文献
This paper presents an experimental characterization of two sources of calcium carbonate, limestone and calcium carbonate precipitate (CCP) used in wet flue gas desulphurization processes. Characterization of the two carbonate sources was carried out by chemical analysis, IR spectra, thermal behavior, particle size distribution for CCP, BET surface area and absorption capacity of SO2 in calcium carbonate suspensions. The absorption temperature, suspension concentration and carbonate grain size were found to be the most influential parameters in the absorption capacity measurements. 相似文献
We give a simpler proof of the well-known result of Matthews and Sumner stating that squares of connected claw-free graphs are vertex-pancyclic. Contrary to the previous proof, our approach does not resort to Fleischner’s result stating that, when restricted to squares of graphs, vertex-pancyclicity and Hamiltonicity are equivalent. The same proof idea already yielded that connected claw-free graphs of even order have a perfect matching, which is another result of Sumner. We conclude by observing that this proof identifies a larger collection of graphs for which the two properties in question hold. 相似文献