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In the development of therapeutic proteins, analytical assessment of structural stability and integrity constitutes an important activity, as protein stability and integrity influence drug efficacy, and ultimately patient safety. Existing analytical methodologies solely rely on relative changes in optical properties such as fluorescence or scattering upon thermal or chemical perturbation. Here, we present an absolute analytical method for assessing protein stability, structure, and unfolding utilizing Taylor dispersion analysis (TDA) and LED-UV fluorescence detection. The developed TDA method measures the change in size (hydrodynamic radius) and intrinsic fluorescence of a protein during in-line denaturation with guanidinium hydrochloride (GuHCl). The conformational stability of the therapeutic antibody adalimumab and human serum albumin were characterized as a function of pH. The simple workflow and low sample consumption (40 ng protein per data point) of the methodology make it ideal for assessing protein characteristics related to stability in early drug development or when having a scarce amount of sample available. 相似文献
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Novel rapid liquid chromatography tandem masspectrometry method for vemurafenib and metabolites in human plasma,including metabolite concentrations at steady state 下载免费PDF全文
Svante Vikingsson Malin Strömqvist Anna Svedberg Johan Hansson Veronica Höiom Henrik Gréen 《Biomedical chromatography : BMC》2016,30(8):1234-1239
A novel, rapid and sensitive liquid chromatography tandem–mass spectrometry method for quantification of vemurafenib in human plasma, that also for the first time allows for metabolite semi‐quantification, was developed and validated to support clinical trials and therapeutic drug monitoring. Vemurafenib was analysed by precipitation with methanol followed by a 1.9 min isocratic liquid chromatography tandem masspectrometry analysis using an Acquity BEH C18 column with methanol and formic acid using isotope labelled internal standards. Analytes were detected in multireaction monitoring mode on a Xevo TQ. Semi‐quantification of vemurafenib metabolites was performed using the same analytical system and sample preparation with gradient elution. The vemurafenib method was successfully validated in the range 0.5–100 μg/mL according to international guidelines. The metabolite method was partially validated owing to the lack of commercially available reference materials. For the first time concentration levels at steady state for melanoma patients treated with vemurafenib is presented. The low abundance of vemurafenib metabolites suggests that they lack clinical significance. Copyright © 2016 John Wiley & Sons, Ltd. 相似文献
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Henrik Høyer Matti Knaapila Jakob Kjelstrup‐Hansen Xuhai Liu Geir Helgesen 《Journal of Polymer Science.Polymer Physics》2012,50(7):477-483
We demonstrate micromechanical strain sensors with integrated readout based on carbon nanocones and discs (CNCs) which are aligned into a string‐like formation using an alternating electric field and studied by AC impedance spectroscopy and electromechanical methods. The CNC particles are first dispersed into a polymer matrix with a particle fraction of 0.1 vol %. This value is well below the percolation threshold (~ 2 vol %), which suppresses particle aggregation and facilitates transparency allowing the use of an UV‐curable polymer. Alignment was carried out with a 1 kHz, 4 kV/cm electric field and is a consequence of dielectrophoretic effect. It develops in minutes and makes the initially insulating, nonaligned material conductive. This is followed by UV curing of the polymer matrix, which renders a solid state device. The stretching of the aligned strings in the cured polymer leads to a reversible piezoresistive effect, and a gauge factor of about 50 is observed. This is in a sharp contrast to CNC films with particle fraction above percolation threshold (13 vol %), which are conductive but not sensitive to stretching. The strings are Ohmic in nature and moreover show higher DC conductivity (22–500 S/m) compared to identically prepared carbon black strings (1–22 S/m). © 2011 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys, 2011 相似文献
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Alexandru Vlasceanu Dr. Søren L. Broman Anne S. Hansen Anders B. Skov Prof. Dr. Martina Cacciarini Prof. Dr. Anders Kadziola Prof. Dr. Henrik G. Kjaergaard Prof. Dr. Kurt V. Mikkelsen Prof. Dr. Mogens Brøndsted Nielsen 《Chemistry (Weinheim an der Bergstrasse, Germany)》2016,22(31):10796-10800
The conversion and efficient storage of solar energy is recognized to hold significant potential with regard to future energy solutions. Molecular solar thermal batteries based on photochromic systems exemplify one possible technology able to harness and apply this potential. Herein is described the synthesis of a macrocycle based on a dimer of the dihydroazulene/vinylheptafulvene (DHA/VHF) photo/thermal couple. By taking advantage of conformational strain, this DHA–DHA macrocycle presents an improved ability to absorb and store incident light energy in chemical bonds (VHF–VHF). A stepwise energy release over two sequential ring‐closing reactions (VHF→DHA) combines the advantages of an initially fast discharge, hypothetically addressing immediate energy consumption needs, followed by a slow process for consistent, long‐term use. This exemplifies another step forward in the molecular engineering and design of functional organic materials towards solar thermal energy storage and release. 相似文献
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Christian Fabiansen Charles W. Yaméogo Sarita Devi Henrik Friis Anura Kurpad Jonathan C. Wells 《Isotopes in environmental and health studies》2017,53(4):344-355
Childhood malnutrition is highly prevalent and associated with high mortality risk. In observational and interventional studies among malnourished children, body composition is increasingly recognised as a key outcome. The deuterium dilution technique has generated high-quality data on body composition in studies of infants and young children in several settings, but its feasibility and accuracy in children suffering from moderate acute malnutrition requires further study. Prior to a large nutritional intervention trial among children with moderate acute malnutrition, we conducted pilot work to develop and adapt the deuterium dilution technique. We refined procedures for administration of isotope doses and collection of saliva. Furthermore, we established that equilibration time in local context is 3?h. These findings and the resulting standard operating procedures are important to improve data quality when using the deuterium dilution technique in malnutrition studies in field conditions, and may encourage a wider use of isotope techniques. 相似文献
70.
Antoine Henrot Henrik Shahgholian 《Transactions of the American Mathematical Society》2002,354(6):2399-2416
Our objective, here, is to generalize our earlier results on the existence of classical convex solution to a free boundary problem with a Bernoulli-type boundary gradient condition and with the -Laplacian as the governing operator. The main theorems of this paper assert that the exterior and the interior free boundary problem with a Bernoulli law, i.e. with a prescribed pressure on the ``free' streamline of the flow, have convex solutions provided the initial domains are convex. The continuous function is subject to certain convexity properties. In our earlier results we have considered the case of constant . In the lines of the proof of the main results we also prove the semi-continuity (up to the boundary) of the gradient of the -capacitary potentials in convex rings, with boundaries.