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101.
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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固体基质室温燐光法(SS-RTP)的检出限依赖于固体基质的RTP背景的高低。本文考察了七种国产滤纸在重原子存在时和不存在时的RTP背景。研究了各种降低滤纸背景的处理方法。研究结果表明,采用NaOH、β-环糊精、EDTA、乙醇和丙酮浸泡或煮沸滤纸,都能不同程度的降低滤纸背景,最大降低率为88%。KI、NaAc、Pb(Ac)_2、TlNO_3和Cscl等重原子可使滤纸背景有较大幅度的增强,而经处理的滤纸,RTP背景较未处理滤纸背景降低一倍以上,最大可降低94.5%。用NaOH溶液处理滤纸,不仅能有效地降低背景,而且能提高分析物的RTP信号。研究表明纤维素伴生物质——木素,可能是产生燐光背景的杂质之一。 相似文献
104.
I. M. J. J. van de Ven-Lucassen M. F. Kemmere P. J. A. M. Kerkhof 《Journal of solution chemistry》1997,26(12):1145-1167
The Taylor dispersion technique is used to measure the ternary mutual diffusion coefficients of aqueous nonelectrolyte solutions at 25°C. The dispersion of the injected solutes is recorded by a differential refractometer and an ultraviolet-visible detector. The diffusion coefficients are calculated directly by fitting the theoretical dispersion equations to about six experimental curves simultaneously. If the ternary diffusion effects in the measured dispersion profiles are not confused by the inaccuracy of the experimental method or an unfavorable relative detector sensitivity, the diffusion coefficients are precise. For the system methanol + acetone + water, it is shown that the Taylor dispersion method is unsuitable for the determination of all the diffusion coefficients if the methanol mole fraction is less than 0.45 or the acetone mole fraction if more than 0.001. 相似文献
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N.C. Dafader M.E. Haque F. Akhtar 《高分子科学》2007,(5):519-523
Natural rubber latex(NRL)and methyl methacrylate(MMA)grafted rubber latex were blended in different ratios and irradiated at various absorbed doses by gamma rays from Co-60 source at room temperature.The tensile properties, swelling ratio and permanent set were measured.The maximum tensile strength and modulus at 500% elongation were obtained at an absorbed dose of 8 kGy.Modulus increases from 6.99 MPa to 9.87 MPa for an increase in proportion of MMA grafted rubber from 40% to 60% in the blend at similar absorbed dose.Elongation at break and swelling ratio decrease with increasing absorbed dose as well as the MMA grafted rubber content in the blends.The decreasing trend of permanent set is high up to 5 kGy absorbed dose,and beyond that dose,it becomes almost flat. 相似文献
107.
通过采用水热晶化与荷电复合分散法制备γ-AlOOH、R-TiO2和α-Al2O3纳米晶体粉末,利用TEM和激光粒度分析仪等手段研究了纳米颗粒水热晶化与荷电复合分散的分散效果及其工艺条件,并探讨了金属氧化物纳米颗粒水热晶化与荷电复合分散机理。结果表明,晶化与荷电复合分散可明显改善金属氧化物纳米颗粒在液相中的分散稳定性,所制得的AlOOH、TiO2和Al2O3纳米颗粒在水中分散放置24 h后的透光率的变化率D24还可分别保持为94.1%、87.7%和82.2%;在水中分散后的粒径分布统计的平均粒径分别为67、70和143 nm。 相似文献
108.
Ashok M. Raichur 《Journal of Dispersion Science and Technology》2013,34(8):1272-1277
Dispersability of colloidal alumina in water has been studied using a rhamnolipid containing biosurfactant. Zeta potential measurements revealed that the surface charge of alumina was altered due to adsorption of the biosurfactant and the iso‐electric‐point of alumina shifted from pH 9 to 6.3. Sedimentation tests indicated that the alumina suspension was completely dispersed for 3–5 hours in the presence of biosurfactant after which some settling was observed. Stability of the suspension in the time period studied was found to be independent of pH. Capillary suction time measurements showed that the alumina suspension was dispersed in the presence of the biosurfactant and varied with pH. Maximum dispersion is obtained in the pH range of 3.5–5 and 7–11 while a minimum is obtained around pH 6. This behavior is consistent with the changes in zeta potential in the presence of the biosurfactant and thus capillary suction time measurements appeared to be more reliable than the sedimentation tests. Optimization studies showed that about 60 mg/g of biosurfactant was necessary for best dispersion and dispersion could be done up to 40% solids. The application of a natural biosurfactant for dispersing colloidal alumina has been demonstrated. 相似文献
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110.