Characterization of the network properties of poly(ethylene glycol)–acrylate hydrogels prepared by variations in the ethanol–water solvent composition during crosslinking copolymerization

The characteristics of poly(ethylene glycol) (PEG)–acrylate hydrogel networks were investigated as a function of the ethanol–water solvent composition during free‐radical crosslinking copolymerization. Macromonomer (88% ω‐methoxy‐PEG–acrylate and 10% ω‐phenoxy‐PEG–acrylate) and crosslinker (2% PEG–diacrylate) concentrations were kept constant. As the copolymerization progressed, the polymer solution in 100% ethanol became increasingly turbid, indicating the development of a heterogeneous network structure. In 100% water, however, the initially turbid polymer solution became increasingly transparent as the crosslinking copolymerization progressed. All the gels were optically clear upon equilibration in water. Kinetic studies, with attenuated total reflectance‐infrared, showed a long induction period, along with a lowered reaction rate, in 100% ethanol, and a decrease in conversion with an increase in ethanol content. These results agree with the UV analysis of the sol fractions, which indicated an increase in the amounts of unreacted PEG–acrylates with an increase in the ethanol content. The gels which were formed with a high ethanol concentration exhibited lower Young's modulus and higher swelling ability, suggesting that the network structure was significantly affected by the solvent composition during free‐radical crosslinking copolymerization. From the stress–strain and swelling experiments, the Flory–Huggins interaction parameter was evaluated. The creep characteristics of the hydrogels were modeled with two Kelvin elements. © 2002 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 40: 2677–2684, 2002     

DFT‐GIAO 1H and 13C NMR prediction of chemical shifts for the configurational assignment of 6β‐hydroxyhyoscyamine diastereoisomers

¹H and ¹³C NMR chemical shift calculations using the density functional theory–gauge including/invariant atomic orbitals (DFT–GIAO) approximation at the B3LYP/6‐311G++(d,p) level of theory have been used to assign both natural diastereoisomers of 6β‐hydroxyhyoscyamine. The theoretical chemical shifts of the ¹H and ¹³C atoms in both isomers were calculated using a previously determined conformational distribution, and the theoretical and experimental values were cross‐compared. For protons, the obtained average absolute differences and root mean square (rms) errors for each comparison showed that the experimental chemical shifts of dextrorotatory and levorotatory 6β‐hydroxyhyoscyamines correlated well with the theoretical values calculated for the (3R,6R,2′S) and (3S,6S,2′S) configurations, respectively, whereas for ¹³C atoms the calculations were unable to differentiate between isomers. The nature of the relatively large chemical shift differences observed in nuclei that share similar chemical environments between isomers was asserted from the same calculations. It is shown that the anisotropic effect of the phenyl group in the tropic ester moiety, positioned under the tropane ring, has a larger shielding effect over one ring side than over the other one. Copyright © 2009 John Wiley & Sons, Ltd.     

Development,validation, and implementation of capillary gel electrophoresis as a replacement for SDS‐PAGE for purity analysis of IgG2 mAbs

Capillary gel electrophoresis (CGE) methods with UV detection were developed for reduced and non‐reduced mAb analysis. These methods can be used to evaluate mAb purity, offering more reproducible quantitation compared with that of traditional SDS‐PAGE methods. These CGE methods have been utilized as platform technology for bioprocess development, formulation development, mAb characterization, drug substance/drug product release testing as well as a required methodology for stability testing. We have found these CGE methods to be applicable across a platform of mAbs in preclinical and clinical development, with the majority of mAbs requiring no modification to the method conditions. This methodology has been ICH validated and transferred to several supporting organizations. The data presented herein describes the development of CGE methodology, platform application to mAb purity analysis, ICH validation, reliability metrics, and considerations on technology enhancement for improved performance and throughput.     

Capillary array isoelectric focusing with laser-induced fluorescence detection: milli-pH unit resolution and yoctomole mass detection limits in a 32-channel system

We report a multiplexed capillary electrophoresis system employing an array of 32 capillaries with a micromachined sheath-flow cuvette as the detection chamber. The sample streams were simultaneously excited with a 473-nm laser beam, and the fluorescence emission was imaged on a CCD camera with a pair of doublet achromat lens. The instrument produced mass detection limits of 380 ± 120 yoctomoles for fluorescein in zone electrophoresis. Capillary isoelectric focusing of fluorescent standards produced peaks with an average width of 0.0029 ± 0.0008 pH. Capillary coating stability limits the reproducibility of the analysis.     

Reactivity of the tethered alkyl uranium bonds of (η5:κ1-C5Me4SiMe2CH2)2U

Uranium-carbon bond reactivity has been investigated with the bis(tethered silylalkyl) uranium metallocene (η⁵:κ¹-C₅Me₄SiMe₂CH₂)₂U, 1. Tert-butyl nitrile, ^tBuCN, inserts into both of the tethered U-C bonds to produce the bis(tethered ketimide) complex [η⁵:κ¹-C₅Me₄SiMe₂CH₂C(^tBu)N]₂U, 2, which has unusually bent U-N-C bond angles. Carbon dioxide also inserts into both U-C bonds of 1 yielding the bis(tethered carboxylate) (C₅Me₄SiMe₂CH₂CO₂)₂U, 3. Neither PhCCPh nor PhCCH insert into the U-C bonds, but PhCCH cleaves the silylalkyl tethers in 1 to generate (C₅Me₄SiMe₃)^1? ligands in the complex (C₅Me₄SiMe₃)₂U(CCPh)₂, 4.     

Chemical-genetic screen identifies riluzole as an enhancer of Wnt/β-catenin signaling in melanoma

To identify new protein and pharmacological regulators of Wnt/β-catenin signaling, we used a cell-based reporter assay to screen a collection of 1857 human-experienced compounds for their ability to enhance activation of the β-catenin reporter by a low concentration of WNT3A. This identified 44 unique compounds, including the FDA-approved drug riluzole, which is presently in clinical trials for treating melanoma. We found that treating melanoma cells with riluzole in?vitro enhances the ability of WNT3A to regulate gene expression, to promote pigmentation, and to decrease cell proliferation. Furthermore riluzole, like WNT3A, decreases metastases in a mouse melanoma model. Interestingly, siRNAs targeting the metabotropic glutamate receptor, GRM1, a reported indirect target of riluzole, enhance β-catenin signaling. The unexpected regulation of β-catenin signaling by both riluzole and GRM1 has implications for the future uses of this drug.     

Development of a capillary gel electrophoresis method for monitoring disulfide isomer heterogeneity in IgG2 antibodies

A CGE method for monitoring the disulfide isomer distribution characteristic of IgG2 MAbs is presented. Disulfide heterogeneity of MAbs has been studied using various chromatographic and electrophoretic methods. Although CGE operates using a different selectivity mechanism from that of sorption chromatographic techniques, similar trends are present in the data, which allow the CGE method to be used as a complementary method for studying disulfide isomer distribution. This article focuses on the optimization of a capillary‐based gel electrophoresis method that can be used to support antibody development including bioprocess optimization, antibody characterization, release, and formulation stability assessment.     

Effect of oceanic turbulence on polarization of stochastic beams

On the basis of the extended Huygens-Fresnel principle and the unified theory of coherence and polarization of light we determine the changes in various polarization properties of stochastic beams propagating through the turbulent clear-water ocean. The ocean-induced fluctuations in the refractive index are described via the recently developed power spectrum which takes into account both temperature and salinity variations. Numerical examples of changes in the spectral density, the degree of polarization and in the polarization ellipse are given for electromagnetic Gaussian Schell-model beams. We demonstrate, in particular, how polarization of the propagating beam is affected by statistical properties of the source and by several parameters of oceanic turbulence. We find that propagation of light beams in the oceanic turbulence resembles that in the atmospheric turbulence qualitatively, however evolution and asymptotic saturation of polarization in the oceanic turbulence occurs at much shorter distances.     

Multiwavelength lasers with homogeneous gain and intensity-dependent loss

Using a simple discrete laser model that incorporates only gain and loss, we showed that lasers with homogeneous gain can have multiwavelength continuous wave output if the loss element is a saturable transmitter. The intensity-dependent loss provides an adaptive balance to the different gain values at different wavelengths. The output power spectrum of the multiwavelength laser will be flat if the laser operates near a peak of the transmission of the intensity-dependent loss.     

Cascading of liquid crystal elastomer photomechanical optical devices

Photomechanical actuation is demonstrated in two coupled liquid crystal elastomer photomechanical optical devices (PODs) acting in series. The response function of an individual POD is characterized and used to predict the temporal response of the coupled system. The predicted coupled-system response agrees with the experiment for several waveforms and frequencies, suggesting that large-scale integration of photomechanical devices is possible.