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1.
固定化牛血清白蛋白吸附材料的制备及其对胆红素的吸附性能 总被引:1,自引:0,他引:1
降低高胆红素血症和重症肝炎患者血液中异常升高的胆红素浓度是血浆交换和血液灌流等疗法的目标之一。本文通过共价键合的方法将牛血清白蛋白(BSA)固定在甲基丙烯酸缩水甘油酯共聚三羟甲基丙烷三甲基丙烯酸酯[poly(GMA-co-TMPTMA)]大孔树脂微球上,制备得到对胆红素具良好吸附性能的固定化BSA吸附材料(BIA),吸附容量达48.7mg/g。由于血清白蛋白对胆红素的强烈相互作用,胆红素溶液中游离BSA的存在会显著降低BIA对胆红素的吸附量。BIA对胆红素的吸附量随吸附温度升高而增加。BIA在-80℃下储存31d后性能仍然稳定,对胆红素的吸附量几乎不变。上述结果表明所制备的BIA为以特异性吸附胆红素为目的的血液净化材料提供了新的选择。 相似文献
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Anna Michnik K. Michalik A. Kluczewska Zofia Drzazga 《Journal of Thermal Analysis and Calorimetry》2006,84(1):113-117
The thermal denaturation process of bovine and human
both fatty acid containing and fatty acid free albumins in aqueous solution
was studied by use of differential scanning calorimetry. Human serum albumins
were found to be more stable than their bovine counterparts. Fatty acid free
albumins were characterized as generally less stable, more susceptible to
aggregation, their unfolding endothermic transition was less cooperative and
with the smaller degree of reversibility. Deconvolution analysis with using
a non-two-state model with two component transitions showed essential differences
in the thermodynamic parameters between all studied albumins, particularly
regarding the high-temperature component transition. 相似文献
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Two classes of binding sites, a single high-affinity site with an association constant of 4·8×106 M−1 and two low-affinity sites with association constant of about 0·05×106 M−1 have been observed in the interaction of Naproxen with bovine serum albumin (BSA). Chemical modification of two tryptophan
residues in BSA with 2-hydroxy-5-nitrobenzyl bromide has led to a reduction in the association constant of the high-affinity
site by 89% and its number of binding sites by 66% suggesting the involvement of tryptophan residues in the high-affinity
site. In contrast, the two low-affinity sites were not affected by the modification. Binding of Naproxen to the low-affinity
sites of BSA induces microdisorganisation of the albumin structure leading to conformational changes as evident from fluorescence
measurements with 1-anilino-8-naphthalenesulphonic acid as the probe. 相似文献
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A calorimetric study of thermal denaturation of bovine serum albumin in aqueous solutions has shown essential differences
in stability of fatty acid containing and defatted albumin. The first one shows a single endotherm peak in DSC curve near
69°C with enthalpy change about 1000 kJ mol-1. Defated albumin melts in two different temperature ranges: near 56 and 69°C with enthalpy changes about 300 and 200 kJ mol-1 respectively. Deconvolution analysis shows that the single endotherm is well approximated as the sum of three independent
two-state transitions. Two transitions of bimodal DSC curve for defatted albumin are not of a two-state type. This molecule
melts probably as two structurally independent parts.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Anna Michnik Katarzyna Michalik Zofia Drzazga 《Journal of Thermal Analysis and Calorimetry》2005,80(2):399-406
Summary The effect of pH on the thermal denaturation of BSA containing fatty acids was studied by use of differential scanning calorimetry (DSC). Thermal scanning of BSA aqueous solutions gave various types of DSC curves depending on the protein concentration and on the pH. The broad bimodal endothermic transition was suggested to be connected with loose protein structure in contradistinction to single peak for compact molecule structure. The propensity toward precipitation at pH conditions ranging from 3.8 to 5 was observed. A scan-rate independent and partly reversible behavior of the thermal heating of BSA was found. Deconvolution of DSC traces in non-two-state model with assumption of two- or three-component transition allowed to study the effect of pH on different parts of BSA molecule. 相似文献
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The interaction between BSA and epicatechin was studied using fluorescence quenching titrations combined with trilinear decomposition method and excitation-emission matrix(EEM)fluorescence.The resolved spectra were highly similar with the actual ones which indicated that the resolved results were reliable.The relevant parameters of the binding process were obtained by quantifying each substance in the complicated mixtures in situ.The quenching was static quenching,epicatechin had a weak interaction with BSA and the binding site was one.The total concentration and the free concentration of quenchers had different effect on the system.The results demonstrated that the method exploited in this article is a useful tool to investigate complicated interactions,avoiding complicated pretreatment and simplify experimental procedure. 相似文献
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采用荧光光谱法和热力学方法研究了3-乙基苯并噻唑螺萘并噁嗪(EBSN)与牛血清白蛋白(BSA)的相互作用.结果表明,在pH为7.46的Tris-HCl及0.01mol·L-1的NaCl介质中,EBSN能强烈猝灭BSA的荧光,猝灭机理为形成复合物的静态猝灭.在298、306和313K时,两者的表观结合常数Kb分别为1.762 0×104,3.396 3×104和6.123 5×104 L·mol-1.与此同时,EBSN与BSA的结合反应是自发的,作用力主要为疏水作用力;BSA和EBSN的工作曲线分别为F0-F=2.439c-8.322和(F0-F)/F=0.061 89c+0.556 6,对BSA和EBSN的检出限分别为0.015mg·L-1和7.61×10-7 mol·L-1. 相似文献
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甲苯胺蓝与牛血清白蛋白作用的光谱特性研究 总被引:1,自引:0,他引:1
应用荧光光谱法研究了甲苯胺蓝(TB)与牛血清白蛋白(BSA)相互作用的光谱特性.测定了TB与BSA在20℃,30℃和40℃3个温度下的结合常数KA分别为8.94×103,1.75×104,1.74×104L/mol,结合位点数n分别为0.66,0.71和0.93.实验表明:TB与BSA的荧光猝灭主要为静态猝灭;利用Gibbs-Helmholtz方程计算得到热力学参数ΔH为49.416kJ,ΔS为244.876J.mol-1.K-1,探讨了它们的相互作用机理,结果表明TB主要以氢键作用力与BSA相互作用;应用同步荧光技术研究了TB对BSA构象的影响. 相似文献
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H. N. Kalpana B. C. Channu Chhabil Dass P. J. Houghton K. N. Thimmaiah 《Journal of Chemical Sciences》2000,112(1):51-61
The interaction of 10-(3′-N-morpholinopropyl)phenoxazine [MPP], 10-(4′-N-morpholinobutyl)phenoxazine [MBP], 10-(3′-N-morpholinopropyl)-2-chlorophenoxazine
[MPCP], 10-(3′-N-piperidinopropyl)-2-chlorophenoxazine [PPCP] or 10-(3′-N-morpholinopropyl)-2-trifluoromethylphenoxazine [MPTP]
with bovine serum albumin (BSA) has been studied by gel filtration and equilibrium dialysis methods. The binding of these
modulators, based on dialysis experiments, has been characterized using the following parameters: percentage of bound drug
(Β), the association constant (K
1), the apparent binding constant (k) and the free energy change (δF‡). The binding of phenoxazine derivatives to serum transporter protein, BSA, is correlated
with their octanol-water partition coefficient, log10
P. In addition, effect of the displacing activities of hydroxyzine and acetylsalicylic acid on the binding of phenoxazine derivatives
to albumin has been studied. Results of the displacement experiments show that phenoxazine benzene rings and tertiary amines
attached to the side chain of the phenoxazine moiety are bound to a hydrophobic area on the albumin molecule. 相似文献
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Campbell W. Robinson Marc H. Siegel Agn s Condemine Conan Fee Thomas Z. Fahidy Bernard R. Glick 《Journal of membrane science》1993,80(1):209-220
A conventional crossflow ultrafiltration (CUF) apparatus was modified by the inclusion of electrodes which permitted a pulsed electric field to be produced across the ultrafiltration membrane (PEF-UF process). Using this apparatus, a discontinuous electrophoretic velocity was imposed upon the proteins being concentrated, opposing their convective movement toward the CUF membrane. This resulted in a lower concentration of rejected solute protein in the fluid boundary layer adjacent to the high-pressure side of the membrane and, hence, in a lower solute-related filtration resistance than in the case of conventional ultrafiltration (zero electric field). Studies of the PEF-UF process with bovine serum albumin (BSA) in the range of 0.5–5% w/v demonstrated a 25–40% decrease in the solute-related resistance to the permeate flux compared to the case of a zero electric field. Accordingly, higher permeate fluxes and, therefore, higher rates of concentration of the protein solution were obtained than for conventional crossflow ultrafiltration. When the electric field was reimposed following a period of operation under conventional CUF conditions, the permeate flux could be restored to nearly the same higher value observed initially for the PEF-UF process. 相似文献
12.
Temperature‐induced conformational transition of bovine serum albumin in neutral aqueous solution by reversed‐phase liquid chromatography 下载免费PDF全文
The temperature‐induced conformational transition of bovine serum albumin (BSA) in neutral aqueous solution was studied using intrinsic fluorescence emission spectrum, reversed‐phase liquid chromatography and sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and the conformation transition thermodynamic parameters were determined in the temperature range 12–50 °C. The results showed that, in the temperature range 12–20 °C, BSA only existed in a single conformation state A, while in the temperature range 22–50 °C, it existed in two different conformation states: A and B. The percentage of conformation state A decreased while that of conformation state B increased with the increase in temperatures, and when temperature approached 50 °C conformation state B accounted for approximately 25% of all conformation states of BSA. In the conformational transition of BSA from conformation state A to conformation state B, the positive enthalpy change, entropy change and free energy changes demonstrated that the conformational transition was endothermic, nonspontaneous and mainly entropy‐driven. © 2013 The Authors. Biomedical Chromatography published by John Wiley & Sons, Ltd. 相似文献
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本文以四乙烯五胺与氯乙酰化聚苯乙烯树脂反应合成PS-TEPA树脂。考察了温度、时间、胺用量以及溶剂等合成条件。研究了PS-TEPA树脂吸附牛血清蛋白的条件,包括吸附温度、吸附时间以及蛋白质的用量。结果发现,当以二氯甲烷作分散剂,合成温度为85℃,反应时间为12 h,投料比为32g四乙烯五胺/g树脂条件下,树脂接枝的增重率和氮元素含量最大;当吸附温度为30℃,吸附时间为12 h,牛血清蛋白溶液的浓度为2.0 mg/mL时,PS-TEPA树脂的蛋白吸附量达到80mg/g树脂,吸附回收率为84%。 相似文献
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采用荧光光谱法研究了水溶液中四溴荧光素、四氯四溴荧光素、二碘荧光素、乙基罗丹明B、健那绿B等5种荧光染料探针与牛血清白蛋白(BSA)的相互作用.实验表明,这5种染料探针同牛血清白蛋白结合时,疏水作用力起决定性作用,静电力起次要作用,相比之下牛血清白蛋白(BSA)结合阴离子的能力最强,其次为中性分子,最后为阳离子.通过疏水作用力,5种染料均是以非极性苯基进入BSA疏水性腔体中同色氨酸残基发生作用,弱氢键的形成加强了这种作用力,且使得光谱间能量转移效率明显提高.5种染料探针的极性部位由于极性和空间效应的原因难以进入腔体内部,致使反应均按接近1∶1的方式进行. 相似文献
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运用荧光光谱、紫外吸收光谱探讨了Fe3+对灯盏花素(BR)与牛血清白蛋白(BSA)相互作用的影响;从Fe3+与BSA的静电作用、配位结合以及Fe3+与BR的配位作用等方面分析了影响BR与BSA相互作用的因素.结果表明,Fe3+不改变BSA与BR的作用机制,但使得二者结合的猝灭常数、结合常数及结合位点数减小. 相似文献