Quantification of Structural Integrity and Stability Using Nanograms of Protein by Flow-Induced Dispersion Analysis

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.     

Iron(III)‐mediated AGET ATRP of styrene using tris(3,6‐dioxaheptyl)amine as a ligand

A commercially available tris(3,6‐dioxaheptyl)amine (TDA‐1) was used as a novel ligand for activator generated by electron transfer atom transfer radical polymerization (AGET ATRP) of styrene in bulk or solution mediated by iron(III) catalyst in the presence of a limited amount of air. FeCl₃ · 6H₂O and (1‐bromoethyl)benzene (PEBr) were used as the catalyst and initiator, respectively; and environmentally benign ascorbic acid (VC) was used as the reducing agent. The polymerizations show the features of “living”/controlled free‐radical polymerizations and well‐defined polystyrenes with molecular weight M_n = 2400–36,500 g/mol and narrow polydispersity (M_w/M_n = 1.11–1.29) were obtained. The “living” feature of the obtained polymer was further confirmed by a chain‐extension experiment. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 2002–2008, 2009     

Luminescent Complexes of Europium (III) with 2-(Phenylethynyl)-1,10-phenanthroline: The Role of the Counterions

New antenna ligand, 2-(phenylethynyl)-1,10-phenanthroline (PEP), and its luminescent Eu (III) complexes, Eu(PEP)₂Cl₃ and Eu(PEP)₂(NO₃)₃, are synthesized and characterized. The synthetic procedure applied is based on reacting of europium salts with ligand in hot acetonitrile solutions in molar ratio 1 to 2. The structure of the complexes is refined by X-ray diffraction based on the single crystals obtained. The compounds [Eu(PEP)₂Cl₃]·2CH₃CN and [Eu(PEP)₂(NO₃)₃]∙2CH₃CN crystalize in monoclinic space group P2_1/n and P2_1/c, respectively, with two acetonitrile solvent molecules. Intra- and inter-ligand π-π stacking interactions are present in solid stat and are realized between the phenanthroline moieties, as well as between the substituents and the phenanthroline units. The optical properties of the complexes are investigated in solid state, acetonitrile and dichloromethane solution. Both compounds exhibit bright red luminescence caused by the organic ligand acting as antenna for sensitization of Eu (III) emission. The newly designed complexes differ in counter ions in the inner coordination sphere, which allows exploring their influence on the stability, molecular and supramolecular structure, fluorescent properties and symmetry of the Eu (III) ion. In addition, molecular simulations are performed in order to explain the observed experimental behavior of the complexes. The discovered structure-properties relationships give insight on the role of the counter ions in the molecular design of new Eu (III) based luminescent materials.     

A Highly Active Iron‐Based Catalyst System for the AGET ATRP of Styrene

The atom transfer radical polymerization of styrene using activators generated by electron transfer (AGET ATRP) has been carried out in bulk in a limited amount of air at 110 °C, using 1,3,5‐(2′‐bromo‐2′‐methylpropionato)benzene (BMPB) as an initiator and FeCl₃ · 6H₂O/tris(3,6‐dioxaheptyl) amine (TDA‐1)/ascorbic acid (VC) as a novel Fe^III‐mediated catalyst system. The results of the polymerizations demonstrate the features of ‘living’/controlled free‐radical polymerization, such as the number‐average molecular weights being close to their corresponding theoretical values and increasing linearly with monomer conversion, and narrow molecular weight distributions ( = 1.18–1.26). The end functionality of the obtained polymers was confirmed by ¹H and ¹³C NMR spectra as well as a chain‐extension reaction.

     

Extractable organic compounds in polyurethane foam with special reference to aromatic amines and derivatives thereof

Methods for determination of aromatic amines and related compounds in flexible toluene diisocyanate (TDI)-based polyurethane (PUR) foam were investigated. The foam was extracted using 0.1% (w/v) aqueous acetic acid (HAc). Extraction solutions were analysed and aromatic amines were determined as ethyl chloroformate (Et) and pentafluoropropionic acid anhydride (PFPA) derivatives. The determinations were performed using liquid chromatography (LC) and mass spectrometry (MS) detection with electrospray ionisation (ESI) or gas chromatography (GC)-MS with chemical ionisation monitoring negative ions (NCI). The Et derivatives were determined using LC-ESI⁺-MS with detection limit of 2 pg of toluenediamine (TDA). The PFPA derivatives were determined using LC-ESI⁻-MS or GC-NCI-MS with detection limits of 0.1 and 0.02 pg of TDA, respectively. Using trideuterium labelled TDA as internal standard, linear calibration curves were obtained in the range of 0.01-0.50 μg ml⁻¹ (n=7), with correlation coefficients >0.999. When plotting calibration curves for TDA-PFPA derivatives determined using LC-MS against TDA-PFPA using GC-MS and TDA-Et using LC-MS, linear curves were obtained. The relative standard deviation (R.S.D.) for determination of TDA in foam extraction solutions were 13%. LC-MS determination of PFPA derivatives was more selective, as compared to LC-MS of Et derivatives.In foam extraction solutions, 2,4- and 2,6-TDA, several isomers of methylenedianiline (MDA) and dimers of TDA/TDI were observed. 2,4-TDA and 4,4′-MDA are possible human carcinogens. Hydrolysis of the extraction solution revealed a large pool of TDA/TDI compounds and oligomers. The concentration of TDA in foam was affected by the extraction media, temperature and duration. The choice of derivatisation procedure also affected the determination of TDA. In extraction solutions from six different commercially available flexible foam qualities 2,4- and 2,6-TDA were found in the range of 0-7 and 0-6 μg g⁻¹ foam, respectively. When flexible foam was heated, considerable higher concentrations of TDA were observed.     

Particle-hole calculations in4He and16O with centre of mass subtracted kinetic energy

It is found that in light nuclei it is very important to use a relative kinetic energy operator. This kinetic energy is mass number dependent and contributes in all odd parity (J*,T) channels. In particular it is seen to produce the spurious state exactly at zero energy in the (1⁻, 0) channel in TDA both in⁴He and¹⁶O. The effect of the relative KE is to decrease the central force attraction and increase the relative importance of the tensor force. The latter is important in RPA. Sussex matrix elements without the hard core are used.     

Supramolecular nanostructures based on bacterial reaction center proteins and quantum dots

Design of the nanostructures based on membrane proteins (the key functional elements of biomembranes) and colloid nanoparticles is a fascinating field at the interface of biochemistry and colloids, nanotechnology and biomedicine. The review discusses the main achievements in the field of ultrathin films prepared from bacterial reaction center proteins and light-harvesting complexes, as well as these complexes tagged with quantum dots. The principles of preparation of these thin films and their structure and properties at different interfaces are described; as well as their characteristics estimated using a combination of the modern interfacial techniques (absorption and fluorescence spectroscopy, atomic force and Brewster angle microscopy, etc.) are discussed. Further approaches to develop the nanostructures based on the membrane proteins and quantum dots are suggested. These supramolecular nanostructures are promising prototypes of the materials for photovoltaic, optoelectronic and biosensing applications.     

核磁共振测井TDA识别油气的影响因素

TDA（时域分析）方法是核磁共振测井的主要方法之一,在判断储层流体类型、性质和孔隙度计算方面有很大的优势,但在应用中却存在一定的局限性. 针对TDA方法在流体识别中存在的一些问题,通过数值模拟不同条件的地层对其在油气识别中的影响因素及适用性进行分析. 研究结果表明：TDA主要用来识别轻烃,采用D9TW采集模式采集核磁自旋回波串,TDA能识别的轻质油粘度<9 mPa.s;长极化时间不变,短极化时间T_WS提高到2 s,TDA能识别的轻质油粘度<4 mPa.s. 双T_W回波串的差（差值信号,转换为孔隙度单位称为差分孔隙度）与地层孔隙度、含烃饱和度、含氢指数、极化函数等影响因素正相关. 对于低信噪比的核磁数据,综合各影响因素,发现烃的差分孔隙度要>1.5 p.u.,TDA方法才能有效识别油气和计算孔隙度.     

SD对壳模型：对结构对原子核集体性质的影响

在SD对壳模型的理论框架下, 利用变分法及TDA近似确定SD对结构. 结果发现利用上述方法确定SD对比以往通过对角化表面δ相互作用来确定SD对结构的方法可以更好地再现原子核低激发谱的集体性质.     

Development of magnetic,ferrite supported palladium catalysts for 2,4-dinitrotoluene hydrogenation

Cobalt, copper, and nickel ferrite spinel nanoparticles have been synthesized by using a combination of sonochemical treatment and combustion. The magnetic nanoparticles have been used as supports to prepare ~4 wt% palladium catalysts. The ferrites were dispersed in an ethanolic solution of Pd(II) nitrate by ultrasonication. The palladium ions were reduced to metallic Pd nanoparticles, which were then attached to the surface of the different metal oxide supports. Thus, three different catalysts (Pd/CoFe₂O₄, Pd/CuFe₂O₄, Pd/NiFe₂O₄) were made and tested in the hydrogenation of 2,4-dinitrotoluene (DNT). A possible reaction mechanism, including the detected species, has been envisaged based on the results. The highest 2,4-diaminotoluene (TDA) yield (99 n/n%) has been achieved by using the Pd/NiFe₂O₄ catalyst. Furthermore, the TDA yield was also reasonable (84.2 n/n%) when the Pd/CoFe₂O₄ catalyst was used. In this case, complete and easy recovery of the catalyst from the reaction medium is ensured, as the ferrite support is fully magnetic. Thus, the catalyst is very well suited for applicationy in the hydrogenation of DNT or other aromatic nitro compounds.