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排序方式: 共有542条查询结果,搜索用时 15 毫秒
181.
Quantitative determinations of ions in a variety of media have been performed traditionally via one of three approaches: optical instrumental methods (e.g., atomic absorption, and inductively-coupled plasma-optical emission or mass spectrometry), “wet” methods, or ion-selective sensors. Each of the approaches, though, possesses limitations including: power/reagent consumption and lack of portability for instrumental techniques; laborious sample-treatment steps for wet methods; and lack of selectivity and sensitivity with sensors when employed with complex samples. Microfluidic device have emerged as a solution to some of these challenges associated with ion analysis. Such systems can integrate multiple sample handling, calibration, and detection steps (“lab-on-a-chip” concept) into a footprint amenable to portability, while requiring small amounts of sample and power. Furthermore, devices can be constructed for multi-analyte detection, either through multiple parallel fluidic architectures or by using arrays of detection elements. This paper reviews recent progress in the development of total-analysis systems for ionic species. Fabrication techniques and various fluid-handling operations are discussed briefly, followed by a number of more mature strategies for microfluidic ion analysis. A variety of approaches expected to comprise the next generation of devices are also presented. 相似文献
182.
Calmodulin-mediated reversible immobilization of enzymes 总被引:1,自引:0,他引:1
Daunert S Bachas LG Schauer-Vukasinovic V Gregory KJ Schrift G Deo S 《Colloids and surfaces. B, Biointerfaces》2007,58(1):20-27
This work demonstrates the use of the protein calmodulin, CaM, as an affinity tag for the reversible immobilization of enzymes on surfaces. Our strategy takes advantage of the of the reversible, calcium-mediated binding of CaM to its ligand phenothiazine and of the ability to produce fusion proteins between CaM and a variety of enzymes to reversibly immobilize enzymes in an oriented fashion to different surfaces. Specifically, we employed two different enzymes, organophosphorus hydrolase (OPH) and beta-lactamase and two different solid supports, a silica surface and cellulose membrane modified by covalently attaching a phenothiazine ligand, to demonstrate the versatility of our immobilization method. Fusion proteins between CaM-OPH and CaM-beta-lactamase were prepared by using genetic engineering strategies to introduce the calmodulin tail at the N-terminus of each of the two enzymes. In the presence of Ca(2+), CaM adopts a conformation that favors interaction between hydrophobic pockets in CaM and phenothiazine, while in the presence of a Ca(2+)-chelating agent such as EGTA, the interaction between CaM and phenothiazine is disrupted, thus allowing for removal of the CaM-fusion protein from the surface under mild conditions. CaM also acts as a spacer molecule, orienting the enzyme away from the surface and toward the solution, which minimizes enzyme interactions with the immobilization surface. Since the method is based on the highly selective binding of CaM to its phenothiazine ligand, and this is covalently immobilized on the surface, the method does not suffer from ligand leaching nor from interference from other proteins present in the cell extract. An additional advantage lies in that the support can be regenerated by passing through EGTA, and then reused for the immobilization of the same or, if desired, a different enzyme. Using a fusion protein approach for immobilization purposes avoids the use of harsh conditions in the immobilization and/or regeneration steps, which could cause inactivation of the immobilized enzyme. Moreover, we have demonstrated that the CaM affinity tag allows immobilization of enzymes on a variety of surfaces without compromising their enzymatic activity substantially; for example, the immobilized OPH retained more than 80% of the activity of the free enzyme. Our results with beta-lactamase showed the feasibility of using a phenothiazine surface in several consecutive loading and regeneration cycles. This can be advantageous when expensive and/or difficult to obtain immobilization surfaces have to be employed; the immobilization surface could be reused to immobilize the same or a different enzyme using the CaM affinity tail. We also determined that the phenothiazine-modified silica particles are stable for long periods of time, i.e., up to 2 years when stored at 4 degrees C. It is envisioned that this type of reversible immobilization may find applications in the development of reversible, reusable biosensors and bioreactors endowed with the additional advantage that the biological element at the surface of the sensor or bioreactor could be replaced under mild conditions when needed to sense or process a different target molecule. 相似文献
183.
Chromatographic sorbents used within the purification of peptide or protein based active pharmaceutical ingredients (APIs) are commonly subjected to caustic regeneration procedures, so-called CIP treatments. While polymeric materials remain unaffected by this treatment, silica-based sorbents are at an intrinsic risk of dissolution under high pH conditions, such as, e.g. 0.1M NaOH. It is common misconception that silica-based materials simply cannot be subjected to alkaline conditions above pH 9. Moreover, most studies covering the chemical stability of HPLC sorbents above pH 9 have been limited to the chromatographic conditions used for the separations themselves. Such studies have used buffered mobile phases up to pH 11 or 12. Very little focus has been put on the stability of the stationary phases when subjected to shorter but harsher pH conditions required for regeneration purposes, such as 0.1M NaOH (pH 13). Knowledge about the amount of so-called leachables, degradation products originating from the stationary phase, is of growing importance for the registration of pharmaceuticals for human use and is addressed in this work. This study compares the chemical stability of different commercially available reversed phase silica materials (C18) that are used in industrial scale preparative HPLC. The silica materials were subjected to NaOH regeneration conditions and it is shown that some materials are able to withstand 0.1M NaOH conditions without significant harm. It is demonstrated that contaminants present in the effluent in the range of 10-50 microg/mL can lead to significant contamination of API product fraction. 相似文献
184.
185.
Cortezzi SS Garcia JS Ferreira CR Braga DP Figueira RC Iaconelli A Souza GH Borges E Eberlin MN 《Analytical and bioanalytical chemistry》2011,401(4):1331-1339
A bottom-up label-free mass spectrometric proteomic strategy was used to analyse the protein profiles of the human embryonic
secretome. Culture media samples used for embryonic culture of patients undergoing intracytoplasmic sperm injection cycles
were selected as a test case for this exploratory proof-of-principle study. The media were stored after embryo transfer and
then pooled into positive (n = 8) and negative (n = 8) implantation groups. The absolute quantitative bottom-up technique employed a multidimensional protein identification
technology based on separation by nano-ultra-high pressure chromatography and identification via tandem nano-electrospray
ionization mass spectrometry with data-independent scanning in a hydrid QqTOF mass spectrometer. By applying quantitative
bottom-up proteomics, unique proteins were found exclusively in both the positive- and negative-implantation groups, which
suggest that competent embryos express and secrete unique biomarker proteins into the surrounding culture medium. The selective
monitoring of these possible secretome biomarkers could make viable procedures using single-embryo transfer. 相似文献
186.
We study a mixed heat and Schrödinger Ginzburg–Landau evolution equation on a bounded two-dimensional domain with an electric current applied on the boundary and a pinning potential term. This is meant to model a superconductor subjected to an applied electric current and electromagnetic field and containing impurities. Such a current is expected to set the vortices in motion, while the pinning term drives them toward minima of the pinning potential and “pins” them there. We derive the limiting dynamics of a finite number of vortices in the limit of a large Ginzburg–Landau parameter, or \({\varepsilon \to 0}\) , when the intensity of the electric current and applied magnetic field on the boundary scale like \({|{\rm log} \, \varepsilon|}\) . We show that the limiting velocity of the vortices is the sum of a Lorentz force, due to the current, and a pinning force. We state an analogous result for a model Ginzburg–Landau equation without magnetic field but with forcing terms. Our proof provides a unified approach to various proofs of dynamics of Ginzburg–Landau vortices. 相似文献
187.
Edkins RM Bettington SL Goeta AE Beeby A 《Dalton transactions (Cambridge, England : 2003)》2011,40(47):12765-12770
The near-infrared two-photon absorption (TPA) spectra of a series of cyclometalated iridium complexes have been measured. These complexes exhibit moderately large TPA cross-sections of approximately 20 GM at the biological relevant wavelength of 800 nm. A new complex has been designed and synthesised, and found to have an increased cross-section of 44 GM at 800 nm. Full photophysical characterisation of this complex is presented. 相似文献
188.
Edkins RM Wriglesworth A Fucke K Bettington SL Beeby A 《Dalton transactions (Cambridge, England : 2003)》2011,40(38):9672-9678
Herein we report the synthesis and photophysical study of tris-heteroleptic complexes of the general formula IrLL'(acac), where L and L' are two differently substituted 2-phenylpyridines (ppyH) and acacH is 2,4-pentanedione, using a combinatorial approach that could be employed for many ligand combinations. The tris-heteroleptic complexes and the analogous bis-heteroleptic complexes of the form IrL(2)(acac) have been studied by a combination of absorption and photoluminescence spectroscopies in conjunction with modelling by DFT and TD-DFT to elucidate the nature and location of the excited state in the novel species. 相似文献
189.
Vanderheiden S Bulat B Zevaco T Jung N Bräse S 《Chemical communications (Cambridge, England)》2011,47(32):9063-9065
A novel access to deuterated and D(3)CO-substituted arenes has been developed using immobilized triazenes as precursors. The linker system and the deuterating cleavage methodology could be shown to be compatible with various functional groups and are therefore suitable for the synthesis of derivatives only hardly available via comparable protocols. 相似文献
190.
Martin CJ Gil B Perera SD Draper SM 《Chemical communications (Cambridge, England)》2011,47(12):3616-3618
With a view to combining the desirable electronic and photochemical properties of hexabenzocoronene (HBC) and the C-C bond forming capabilities of thiophenes, 1-(3-thienyl)-2,3,4,5,6-penta(4-tert-butyl-phenyl)benzene (1) was oxidised using FeCl(3). The resulting products, superaromatic thiophene (2) and its 5,5'-dimer (3), are S-HBC systems and provide a new pair of spectral comparators. 相似文献