木瓜蛋白酶在染料Cibacron Blue F3GA纤维素亲和膜上的吸附研究

以纤维素滤纸膜为载体,染料Cibacron Blue F3GA为配基,制备了一种新型亲和膜色谱介质。采用扫描电镜、红外光谱、元素分析等方法对亲和膜介质进行鉴定与表征,该膜具有良好的色谱性能。亲和膜对F3GA的键合质量摩尔浓度达93.7μmol/g。研究了木瓜蛋白酶在亲和膜上的吸附行为,实验表明:在30℃下、酶质量浓度为2 mg/mL、pH=8.0时,吸附质量比可达57.9 mg/g,改变pH值及离子强度等条件对吸附质量比有明显的影响。在最适条件下吸附遵循Langmuir型吸附。可以初步推断,纤维素滤纸膜可以制成性能优良的亲和膜色谱介质,成本低廉,适合工业化分离纯化生物大分子。

Affinity Adsorption of Bilirubin on Cibacron Blue F3GA-modified Microporous Polytetrafluoroethylene Capillary

Bilirubin removal from human plasma was obtained via an affinity microporous polytetrafluoroethylene（MPTFE） capillary. The new adsorbent comprised grafted glycidyl methacrylate（GMA） via radiation-induced polymerization as hydrophilic coating and reactive sites; ethylenediamine（EDA） as a spacer arm; Cibacron Blue F3GA（CB F3GA） as an affinity ligand; MPTFE capillary as the supporting matrix. The average density of CB F3GA attachment to MPTFE capillaries was found to be 136.5 μmol/g. The capacity of bilirubin adsorbed on affinity MPTFE capillaries is 76.1 mg bilirubin/g polymer（at 25℃）. This new adsorbent has advantages over both membrane and traditional micro-column, and this system is stable and easy to operate. The results of blood tests suggest the CB F3GA affinity capillary has good blood compatibility.

Preparation of Cibacron Blue F3GA Bonded Poly (styrenedivinylbenzene) (PSDVB) Microbeads Used for High Performance Affinity Chromatography

Rigid PSDVB microbeads have been modified with poly(vinyl alcohol) (PVA) adsorbed on their surface to produce an affinity medium. Then Cibacron Blue F3GA was covalently attached to the supports. The initial concentration of PVA has effect on the adsorption of PVA. The non-specific interaction of bovine serum albumin (BSA) on the microbeads decreases with the PVA adsorbed. The pH stability test shows that the affinity medium is stable up to pH 11.0. And it has specific interaction with lysozyme, but not with pepsin.

三嗪染料Cibacron Blue F3 GA-吐温80亲和分离介质的合成与应用

研究了一种新的亲和分离介质三嗪染料CibacronBlueF3GA修饰吐温 80的合成条件。采用正交设计合成了染料 吐温 80。最佳反应条件为 :吐温 80 ,4 0 5mmol/L ;CibacronBlueF3GA ,1 0 1mmol/L ;氢氧化钠 ,2 0mol/L ;油浴温度 ,110℃ ;反应时间 ,4h。染料转化率 72 3 %。用溶剂萃取法分离出染料 吐温 80 ,并做了紫外测定。应用该合成物在吐温 80 盐液 固萃取体系中 ,对猪心肌匀浆中乳酸脱氢酶的亲和分离纯化做了研究。结果表明 ,在硫酸铵体系中 ,一步萃取酶活收得率 90 % ,纯化倍数 4 5 ;在磷酸钾盐体系中 ,一步萃取酶活收得率 95 % ,纯化倍数 6 0。

用Blue Sepharose CL-6B快速纯化天花粉蛋白

差光谱显示染料cibacronblueF3GA与天花粉蛋白 (TCS)有特异性结合 ,复合物在可见光部分的最大吸收波长在 690nm ,摩尔消光系数ε =2 6× 1 0 - 3 (mol/L) - 1·cm- 1,解离常数Kd =1 8μmol/L,0 5mol/LNaCl可使复合物解离 .根据这一特点 ,用Blue SepharoseCL 6B凝胶从栝篓块茎中亲和纯化了TCS .此法快速、简便、高效 ,易于大量制备 .

活性蓝F3GA固载的壳聚糖微球对过氧化氢酶的吸附

将反相悬浮交联法制备的壳聚糖微球树脂与三嗪染料活性蓝F3GA(Cibacron Blue F3GA)反应,制得亲和色谱填料,并采用扫描电镜、红外光谱、染料泄漏率测定等对此填料进行鉴定与表征.该填料具有良好的色谱性能,在pH为7.0的磷酸盐缓冲液体系下,对过氧化氢酶的饱和吸附容量为24.6 mg/g,改变pH值及离子强度对结合量有明显影响.

汽巴蓝F3GA染料亲和色谱介质的制备及其对黑芸豆凝集素的特异性吸附

采用反相悬浮再生法制备6 g/100 mL的琼脂糖微球,采用环氧氯丙烷对琼脂糖微球进行二次交联,并进行染料配体汽巴蓝F3GA的偶联,得到一种新型活性Blue Beads 6FF微球。通过傅里叶变换红外光谱、扫描电子显微镜、压力流速曲线、溶胀率、泄漏率和孔隙率等对微球物理性能进行表征。结果发现,微球球形度较好,粒径大小均一,平均粒径为56.37μm左右,具有较高的孔隙率、溶胀率及广范围的pH值稳定性,且耐压能力强。以黑芸豆凝集素为目标蛋白,通过Langmuir吸附等温式拟合,结果表明该填料对黑芸豆蛋白有较高的吸附量(19.7 mg/mL),微球吸附性能良好,可实现对凝集素蛋白的快速分离和纯化。

聚丙烯膜接枝改性亲和膜的制备与表征

采用预辐射接枝法在聚丙烯基膜(PP)上接枝甲基丙烯酸缩水甘油酯(GMA),再将配基辛巴蓝(CBD)固载于GMA聚合链上制备了一种亲和膜。考察了预辐射剂量、接枝单体浓度、反应时间和温度、Mohr’s盐浓度对GMA聚合接枝的影响,获得了较佳的条件:GMA浓度20%(v/v),反应温度70℃,反应时间3h,Mohr’s盐浓度0.01%(wt),CBD溶液浓度25mg·mL-1。用红外光谱(FTIR)和扫描电镜(SEM)分析和观察了该亲和膜的表面形貌特征;在37℃和60mg·L-1的胆红素溶液中对该亲和膜进行静态吸附平衡实验,结果表明,经2.5h吸附达到平衡,其吸附容量可达50mg·g-1。

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

A series of aminated chitosan microspheres were prepared from chitosan,and then an affinity dye-ligand,Cibacron Blue F3GA,was covalently coupled.The maximum attachment of cibacron blue F3GA was 494 μmol/g.Albumin adsorption onto cibacron blue F3GA-attached chitosan microspheres was investigated.The human serum albumin (HSA) equilibrium adsorption capacity was 79.8 mg/g (wet beads).The adsorption equilibrium isotherms appeared to follow a typical Langmuir isotherm.The highest desorption ratio (100%) was achieved by using an eluent of 1.0 mol/L NaSCN (pH 8.0).The cibacron blue F3GA-attached chitosan microspheres could be reused without significant decreases in the adsorption capacities.

尼龙亲和膜的制备及其对木瓜蛋白酶的分离纯化研究

以尼龙膜为基质,键合壳聚糖的方法对尼龙膜进行改性,使膜的非特异性吸附大大降低,并偶联活性染料亲和基Cibacron Blue F3GA,得到一种新的染料亲和膜.该亲和膜具有良好的色谱性能,对木瓜蛋白酶有较高的吸附量(235.3 mg/g),吸附行为满足Freundlich吸附模型,pH值和离子强度对吸附量有明显影响.使用该亲和膜对木瓜粉中的木瓜蛋白酶进行分离纯化,纯化倍数可达46.5倍.

开管毛细管亲和液相色谱初探

提出了一种新的亲和色谱模式开管毛细管亲和液相色谱。在内径为 5 0 μm的毛细管内表面键合一层三嗪染料配体 ,利用毛细管电泳仪的压力系统进行亲和色谱实验。采用流动相切换洗脱技术 ,牛血清白蛋白和溶菌酶获得了有效的分离。连续运行 10次 ,溶菌酶的出峰时间、峰面积和峰高的相对标准偏差分别为0 1% ,4 3% ,3 7%。在 8 6ng～ 2 8 7ng范围内 ,溶菌酶的进样量与峰面积和峰高都呈线性关系 ,其相关系数分别为 0 9946和 0

染料结合2μm无孔硅胶亲和填料的制备与应用

在 ２ μｍ 无孔硅胶表面键合 ３氨丙基三乙氧基硅烷（ Ａ Ｐ Ｓ），并与三嗪染料活性蓝 Ｆ３ Ｇ Ａ（ Ｃｉｂａｃｒｏｎ Ｂｌｕｅ Ｆ３ Ｇ Ａ， Ｃ Ｂ）反应，制得亲和色谱填料，并采用扫描电镜、元素分析、ｐ Ｈ 稳定性测试对此填料进行鉴定与表征。该填料具有良好的色谱性能，且对生物大分子有一定的亲和选择性，改变 ｐ Ｈ 值及离子强度对溶菌酶的结合量有明显影响，可用于分离卵清蛋白（ Ｏｖａｌ）和溶菌酶（ Ｌｙｓ），且对 α，β，γ球蛋白有不同的亲和作用，并可从鸡蛋清中制备少量溶菌酶。

Adsorption of Human Serum Albumin onto PVA-coated Affinity Microporous PTFE Capillary

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.

白蛋白在以染料为配基的醋酸纤维滤棒上的等温吸附行为(英文)

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。