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1.
采用溶胶-凝胶法制备无孔脲醛树脂-二氧化锆复合微球,在碳酸盐缓冲溶液中,利用微球表面的酰胺键与三嗪染料-活性艳蓝X-BR分子中三嗪环上的活泼氯反应,将染料键合到微球表面,以牛血清白蛋白(BSA)为样品,考察了不同条件下,该萃取剂对BSA的萃取能力。结果表明,该萃取剂对BSA的饱和萃取量达到30.3 mg.g-1,同时萃取的蛋白易于洗脱,1.0 mol.L-1氰化钾的洗脱率达到93.5%。  相似文献   

2.
李京华  王俊德  刘学良 《色谱》2002,20(5):419-422
 以醋酸纤维滤棒为基质 ,染料CibacronBlueF3GA为配基 ,合成了一种新的染料亲和介质 ;分别以牛血清白蛋白 (BSA)和人血清白蛋白 (HSA)为对象 ,用静态法进行了吸附实验 ,得到了相应的亲和等温吸附曲线 ;对曲线按Langmuir模型和Freundlich模型分别进行拟合 ;结果表明 ,醋酸纤维滤棒染料亲和介质对BSA和HSA的等温吸附遵循Freundlich模型。采用该亲和介质装柱并分离实际样品人血浆 ,可得到纯化的人血清白蛋白。  相似文献   

3.
壳聚糖亲和磁性毫微粒的制备及其对蛋白质的吸附性能研究   总被引:35,自引:0,他引:35  
以壳聚糖为包裹材料包埋自制的磁流体 ,制备了具有核 壳结构的磁性毫微粒 ,并偶联色素配基CibacronBlue 3GA(偶联量 1 4 .5μmol/mL)得到了一种新型亲和磁性毫微粒 .结果表明 ,所得亲和磁性微球具有较窄的粒径分布、形状规整 .以牛血清白蛋白 (BSA)和溶菌酶 (Lys)为目标蛋白 ,考察了该亲和磁性毫微粒的吸附性能 ,发现其对BSA和Lys的吸附量分别为 4和 2 8mg/g,吸附行为满足Langmuir吸附等温式 ,且对时间依赖性小而对溶液离子强度敏感 .  相似文献   

4.
采用模板法制备的单分散磁性硅胶微球,经过表面修饰偶联上亚氨基二乙酸(IDA),与过渡金属离子Cu2 螯合,制成一种新型的磁性固定化金属亲和纯化载体。用牛血清白蛋白(BSA)作为模型进行磁性固定化金属亲和吸附蛋白的研究,结果表明,BSA在磁性亲和载体上的吸附可用Langmuir吸附方程描述,对BSA的饱和吸附量为90mg/g。将磁性亲和载体用于带有组氨酸标签的镇痛抗肿瘤多肽(analgesic-antitumorpeptide,AGAP)纯化,在未经过滤的细胞裂解液中可以将AGAP一步纯化,非特异性吸附低,操作简便,完全适用于含有组氨酸标记的重组多肽或蛋白的分离纯化。  相似文献   

5.
丁玲  李曦  张超灿 《化学研究》2010,21(1):19-22
以纳米级四氧化三铁为磁性载体,以苯乙烯为单体,用微悬浮聚合法制备了聚苯乙烯磁性微球;以牛血清白蛋白(BSA)为模型蛋白,用荧光光谱仪和紫外-可见吸收光谱仪研究了磁性微球与BSA的相互作用.结果表明,磁性微球与BSA结合反应的猝灭机理为静态猝灭.  相似文献   

6.
胡新良  王勤  王宇 《广州化学》2013,(4):46-49,21
壳聚糖(CS)分子中含有游离态的氨基和羟基,是一种高效的离子吸附剂,而且氨基可质子化形成阳离子,使得壳聚糖对阴离子及两性化合物都具有较强的吸附能力.壳聚糖引入羧甲基后水溶性和反应活性大大增强.实验中以乳化-化学交联法制备壳聚糖/羧甲基壳聚糖(CMC)四种共混微球,通过对比,发现当CS与CMC的质量比为4∶1、2∶3时,成球硬度好,形状明显,大小均匀.随着CMC含量的增加,所成微球对牛血清白蛋白(BSA)的吸附能力逐渐增强.因此,当CS与CMC质量比为2∶3时共混微球自身的物理形态与吸附性能均达到良好状态,是共混高效离子吸附剂的最佳比例选择.  相似文献   

7.
以微米级的P(AM-co-10%MAA)共聚物为模板,在反相悬浮体系中,通过外源沉积的方法合成了硫化镉复合微球CdS-P(AM-co-10%MAA)。以牛血清白蛋白为主要目标蛋白,研究微球对蛋白质的吸附性能。结果表明,牛血清白蛋白在CdS复合微球上的最大吸附量为35.84mg/g,其吸附模式符合Langmuir吸附定律。在20~50℃范围内,随着温度的升高,复合微球对蛋白质的吸附量增大。在磷酸缓冲溶液体系中,CdS-P(AM-co-10%MAA)复合微球对蛋白质的吸附量在蛋白质等电点时达到最大;此外,复合微球不仅对BSA具有一定的吸附能力,对其它蛋白质同样具有一定的吸附能力,说明该微球具有开发为分离材料的潜力。  相似文献   

8.
磁性壳聚糖-聚丙烯酸微球的制备及表征   总被引:12,自引:0,他引:12  
罗志敏  马秀玲  陈盛  钱伟 《化学通报》2005,68(7):551-554
壳聚糖通过与丙烯酸接枝共聚制得壳聚糖聚丙烯酸悬浮液,在铁磁流体(Fe3O4)与聚乙二醇(分散剂)存在下通过与戊二醛交联,制备了磁性壳聚糖聚丙烯酸微球。用扫描电镜、红外光谱对合成的高分子微球进行形貌观察和结构表征,并进行了元素分析和磁性能测试,研究了磁性微球对牛血清白蛋白(BSA)的吸附效果。结果表明,合成的磁性微球外表呈球形,粒径为100~400nm;当Fe含量为2.47%时,磁性微球的饱和磁化强度约为1.30emug,磁矫顽力为280Oe,磁化率为2.16×10-4(常温下),属于顺磁性材料;其对BSA有较好的吸附效果,饱和吸附量约为400mgg。  相似文献   

9.
采用荧光和紫外-可见吸收光谱,研究了染料木素与牛血清白蛋白(BSA)的相互作用.结果表明染料木素对BSA有较强的荧光猝灭作用;根据Stern-Volmer方程得到染料木素与BSA之间的结合常数KA为4.37×106(27 ℃)、6.45×10b(37℃)和6.76×106(47℃).根据Forster非辐射能量转移理论,求出了染料木素与BSA之间的结合距离为2.64 nm(27℃)、2.68mm(37℃)和2.71 nm(47℃).热力学数据表明该药物与牛血清白蛋白的相互作用是一个吉布斯自由能降低的自发过程,且二者之间的主要作用力类型为静电引力,同时用同步荧光光谱探讨了染料木素对BSA构象的影响.  相似文献   

10.
以亲水性乙烯-乙烯醇共聚物(EVAL)为膜吸附剂基质材料,阳离子交换树脂D061为功能树脂,采用干-湿法纺丝技术,制备了树脂填充EVAL中空纤维膜吸附剂。实验采用连续循环膜过滤工艺,研究了这种吸附剂对目标蛋白(牛血清白蛋白-BSA)的吸附和脱附性能。考察了树脂填充量、缓冲溶液pH值等对蛋白质吸附和脱附性能的影响。实验结果表明,树脂填充EVAL中空纤维膜吸附剂对目标蛋白BSA具有较高的吸附容量和脱附率,随着树脂填充量的增加,膜吸附剂对BSA吸附容量增加,并且最大吸附容量在牛血清蛋白等电点(4.8~5.2)附近出现。当树脂填充量65%,缓冲溶液pH4.5时树脂填充EVAL中空纤维膜吸附剂对BSA的吸附容量可达到71.47mgBSA/g膜吸附剂。  相似文献   

11.
染料壳聚糖微球的制备及其对人血清白蛋白吸附性能研究   总被引:1,自引:0,他引:1  
人血清白蛋白(Human Serum Albumin,HSA)是血浆中含量最丰富的蛋白质,约占血浆总蛋白的60%.在人体内,HSA有许多重要的生理功能[1],临床上广泛应用于手术输血和危重病人补液,治疗创伤休克、烧伤、水肿和低白蛋白血症等,而且能增强人体抵抗能力,是迄今为止产量最大、临床用量最大  相似文献   

12.
Cibacron Blue F3GA was immobilized on poly(hydroxyethyl methacrylate) cryogel and it was used for selective and efficient depletion of albumin from human serum. The poly(hydroxyethyl methacrylate) was selected as the basic component because of its inertness, mechanical strength, chemical and biological stability, and biocompatibility. Cibacron Blue F3GA was covalently attached to the poly(hydroxyethyl methacrylate) cryogel to produce poly(hydroxyethyl methacrylate)-Cibacron Blue F3GA cryogel affinity column. The poly(hydroxyethyl methacrylate)-Cibacron Blue F3GA cryogel was characterized with respect to gelation yield, swelling degree, total volume of macropores, Fourier Transform Infrared spectroscopy, and scanning electron microscopy. It was found that the maximum amount of adsorption (343 mg/g of dry cryogel) obtained from experimental results is very close to the calculated Langmuir adsorption capacity (345 mg/g of dry cryogel). The maximum adsorption capacity for poly(hydroxyethyl methacrylate)-Cibacron Blue F3GA cryogel column was obtained as 950 mg/g of dry cryogel for nondiluted serum. The adsorption capacity decreased with increasing dilution ratios while the depletion ratio of albumin remained as 77% in serum sample. Finally, the poly(hydroxyethyl methacrylate)-Cibacron Blue F3GA cryogel was optimized for using in the fast protein liquid chromatography system for rapid removal of the high abundant proteins from the human serum.  相似文献   

13.
Affinity dye-ligand Cibacron Blue F3GA(CB F3GA) was covalently coupled with poly(vinyl alcohol)(PVA) coated on the inner surface of microporous poly(tetra-fluoroethylene)(MPTFE) membranous capillary. The PVA-coated PTFE capillary surface was characterized by XPS and FESEM. The grafting degree of PVA and the amount of CB F3GA immobilized onto the membranous capillary were 23.5 mg/g and 89.6 pmol/g, respectively. These dyed membranous capillaries were chemically and mechanically stable, and could be reproducibly prepared. Human serum albumin(HSA) was selected as model protein. The saturation adsorbance of the dye attached membranous capillary was 85.3 mg HSA/g, while the capacity of non-specific adsorption for HSA was less than 0.3 mg/g.  相似文献   

14.
由大分子单体法制得了聚醋酸乙烯酯接枝聚苯乙烯(PVAc-g-PSt)微球,使该微球在碱性条件下醇解形成了聚乙烯醇接枝聚苯乙烯(PVA-g-PSt)微球.经X射线光电子能谱对PVA-g-PSt微球表层组成的表征,发现微球具有以PVA为壳、PSt为核的核壳结构.进而利用汽巴蓝F3GA(CB)与PVA-g-PSt微球进行亲核反应,制得了CB功能化的PVA-g-PSt微球,由元素分析定量出了固定在微球表面的CB含量.用透射电子显微镜对微球的粒径与形态进行了表征,发现微球表面在CB功能化后变得相对粗糙,粒径大小基本保持不变,但形态更加规整.比较了3种不同微球对蛋白质的吸附,考察了吸附动力学和影响吸附的因素,发现起始牛血清蛋白(BSA)浓度、pH值、离子强度对微球吸附BSA有明显的影响,并利用Zeta电位探讨了微球与蛋白质间的相互作用机理.最后用硫氰酸钠(NaSCN)进行解吸,计算解吸率最高可达到95.7%,该CB功能化微球可以重复利用,吸附率仅减少5.3%.  相似文献   

15.
Commercially available microporous polyamide hollow fibres are modified by acid hydrolysis to activate the reactive groups and subsequently binding of the ligand, i.e. Cibacron Blue F3GA. Then the Cibacron Blue F3GA-derived hollow fibres were loaded with different metal ions (i.e. Zn(II), Cu(II), Ni(II)) to form the metal chelate. The internal polymer matrix was characterised by scanning electron microscopy. The effects of pH, initial concentration of lysozyme, metal type and temperature on the adsorption of lysozyme to the metal–chelated hollow fibres were examined in a batch reactor. The non-specific adsorption of lysozyme onto the polyamide hollow fibres was 1.8 mg/g. Cibacron Blue F3GA immobilisation increased the lysozyme adsorption up to 62.3 mg/g. Metal–chelated hollow fibres showed a significant increase of the adsorption efficiency. Lysozyme adsorption capacities of Zn(II), Cu(II) and Ni(II)-chelated hollow fibres were different. The maximum capacities of Zn(II), Cu(II) or Ni(II)-chelated hollow fibres were 144.2, 75.2 and 68.6 mg/g, respectively. Significant amount of the adsorbed lysozyme (up to 97%) was eluted in 1 h in the elution medium containing 1.0 M NaSCN at pH 8.0 and 25 mM EDTA at pH 4.9. Repeated adsorption–desorption process showed that this novel metal–chelated polyamide hollow fibres are suitable for lysozyme adsorption.  相似文献   

16.
Interferons are potent biologically active proteins synthesized and secreted by somatic cells of all mammalian species. Dye-affinity adsorption is increasingly used for protein separation. Hollow fibers have several advantages as adsorbents in comparison to conventional bead supports because they are not compressible and they eliminate internal diffusion limitations. The aim of this study was to explore in detail the performance of polyamide hollow fibers to which Cibacron Blue F3GA was attached for adsorption of recombinant interferon-α (rHuIFN-α). The hollow fiber was characterized by scanning electron microscopy. These dye-carrying hollow fibers (35.8 μmol/g) were used in the rHuIFN-α adsorption-elution studies. The effects of initial concentration of rHuIFN-α, medium pH, ionic strength and temperature on the adsorption efficiency of dye-attached hollow fibers were studied in a batch system. The non-specific adsorption of rHuIFN-α on the hollow fibers was 1.2 mg/g. Cibacron Blue F3GA attachment significantly increased the rHuIFN-α adsorption up to 99.8 mg/g. Significant amount of the adsorbed rHuIFN-α (up to 94.8%) was eluted in 1 h in the elution medium containing 1.0 M NaCl. In order to determine the effects of adsorption conditions on possible conformational changes of rHuIFN-α structure, fluorescence spectrophotometry was employed. We resulted that dye-affinity hollow fibers can be applied for rHuIFN-α adsorption without causing any significant conformational changes. Repeated adsorption/elution processes showed that these dye-attached hollow fibers are suitable for rHuIFN-α adsorption.  相似文献   

17.
利用j嗪染料辛巴蓝F-3GA修饰经戊二醛交联的啤酒废酵母菌,得到一种新型染料亲和吸附剂.辛巴蓝F-3GA的固载量为161.1 mg/g.以溶菌酶为研究对象,考察吸附时间、酶初始浓度、pH值、离子强度等因素对吸附率的影响.结果表明:当pH=7.0时,其对溶菌酶有较高的吸附量(229.1 mg/g),吸附性能明显优于未接枝...  相似文献   

18.
卜春苗  王有贤  龚波林  阎超 《色谱》2008,26(3):378-383
以3.0 μm无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯树脂为基质,与三嗪染料活性蓝F3GA(Cibacron Blue F3GA)反应,制备出 一种固定化染料聚合物高效亲和色谱填料。考察了应用该填料时流动相中的盐离子浓度、有机溶剂及流速等对牛血清白蛋白(BSA)和 溶菌酶(Lys)保留行为的影响。实验结果表明,该染料亲和填料具有良好的色谱性能。利用前沿色谱法测定了染料与溶菌酶之间的表观 解离常数为5.26×10-5 mol/L。使用该填料成功地从鸡蛋清和小牛血清中分别分离纯化了Lys和BSA,十二烷基硫酸钠-聚丙烯酰胺凝胶 电泳(SDS-PAGE)分析显示Lys和BSA的纯度分别为95%和92%。  相似文献   

19.
A porous silica of nominal 5 microns particle diameter and 30 nm pore size (Nucleosil 300-5) and a non-porous silica of nominal 1.5 microns particle diameter were activated with 3-mercaptopropyltriethoxysilane (MPTS), followed by the immobilization of the triazine dye, Cibacron Blue F3GA. Various biomimetic dye sorbents with graduated ligand densities between 1 mumol/m2 and 0.01 mumol/m2 were prepared. The capacities and the association constants associated with the binding of lysozyme to these sorbents were determined by frontal analysis experiments [J. Chromatogr., 476 (1989) 205-225]. Due to the ability of the Cibacron Blue F3GA-modified silicas to act as mixed mode coulombic and hydrophobic interaction sorbents and the highly charged nature of the surface structure of lysozyme (pl 11), two mobile phase conditions were examined. In one case a 0.1 M phosphate buffer, pH 7.8, was used as the equilibration and loading buffer, in the second case 1 M sodium chloride-0.1 M phosphate buffer, pH 7.8 was employed as the equilibration and loading buffer to monitor the influence of ionic interactions. The elution was performed in each case with a 2.5 M potassium thiocyanate solution. With the porous silica dye sorbents and 1 M NaCl present in the loading buffer, the highest capacity was achieved when Cibacron Blue F3GA was immobilised to the level of 0.1 mumol/m2. In the case of the non-porous silica dye sorbents, the maximum protein capacity was achieved when 0.5 mumol/m2 dye were immobilised onto the support. Evaluation of the frontal breakthrough curves confirmed that the kinetics of adsorption of lysozyme onto the non-porous sorbent were substantially faster than the adsorption of lysozyme onto the porous sorbent due to the absence of pore diffusion effects in case of the non-porous support. Furthermore, the adsorption of lysozyme on both sorbents was faster when no salt was added to the loading buffer, indicating that there is either conformational or reorientation effects operating during the specific binding of the protein to the dye ligand, or that the interaction is proceeding through the participation of a second class of binding sites. The magnitude of the association constants, Ka, for the lysozyme-Cibacron Blue F3GA systems were found to be dependent on the ligand density of the sorbent. With decreasing ligand density, the protein-ligand interaction became stronger, e.g. Ka values became larger. These results confirm earlier observations on the effect of ligand steric compression on the affinate-ligand association constant, e.g. the protein needs sufficient space to interact with the ligand in an optimum way.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

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