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。

Cibacron Blue亲和层析及应用

以Ｆ３ＧＡ（Ｃｉｂａｃｒｏｎ Ｂｌｕｅ Ｆ３ＧＡ）为配基建立了一种可用于免疫毒素（ＩＴ）分离纯化的亲和层析方法。实验中用三种不同来源的核糖体灭活蛋白（ＲＩＰ），即蓖麻毒素Ａ链（ＲＴＡ），苦瓜毒素（ｍｏｍｏｒｄｉｎ，ＭＴ）和Ｓａｐｏｒｉｎ，以探讨ＲＩＰ与Ｆ３ＧＡ的相互作用。分析显示三种ＲＩＰ均能引起Ｆ３ＧＡ吸收光诸明显红移，提示ＲＩＰ均可与Ｆ３ＧＡ发生特异结合。将Ｆ３ＧＡ与Ｓｅｐｈａｄｅｘ交联可获得Ｂｌｕｅｄｅｘ。Ｂｌｕｅｄｅｘ亲和层析是一种经济有效，简单易行，便于在各类实验室中使用的蛋白质亲和层析技术。结果表明：在低盐溶液中ＲＴＡ和ＭＴ均可迅速地与Ｂｌｕｅｄｅｘ结合，而在高盐溶液中（０．６５ｍｏｌ／ＬＮａＣｌ）又极易被洗脱回收。这一技术用于免疫毒素的研究可有效地去除游离抗体，而不影响其杀伤活性。

大豆蛋白纤维/竹纤维混纺针织物的Cibacron FN染料染色

介绍了CibacronFN染料的结构及染色性能,重点探讨了CibacronFN染料染大豆蛋白纤维/竹纤维混纺针织物的条件,染色温度为70℃,保温时间为60min,染浅色时Na2SO4的用量为8～15g/L,染中色时为15～50g/L,染浅色时Na2CO3的用量为5～10g/L,染中色时为13～15g/L。确定了Cibacron黄FN2R、Cibacron红FN R、Cibacron蓝FN R做三元色,对染色中存在的问题进行了分析,找出了原因,给出了解决办法。

Cibacron LS染料低盐染色工艺

针对 Cibacron LS 在30g/L 的低盐染色条件下,对影响染色 K/S 值的染色温度、上染时间、固色时间、纯碱浓度、染色浴比等诸多工艺参数逐一进行试验分析,得出了 Cibacron LS 低盐染色的最佳工艺。

Cibacron Red H的结构剖析

本文利用HNMR、MS、PC、UV-Vis方法剖析了Cibacron Red H的结构,并合成了该染料加以验证。

CibacronFN染料在大豆纤维/天丝混纺织物上的应用

介绍了CibacronFN染料的结构及染色性能,重点探讨了CibacronFN染料染大豆纤维/天丝混纺织物的条件:染色温度为70℃,保温时间为60 min,染浅色时Na2SO4的用量为8-15 g/L,染中色时为15-50 g/L, 染浅色时Na2CO3的用量为5-10 g/L,染中色时为13-15 g/L。确定了CibacronFN2R黄、CibacronFNR红、CibacronFNR蓝做三元色,对染色中存在的问题进行了分析,找出了原因,给出了解决办法。

用Cibacron FN染料染大豆/竹纤维混纺织物的工艺探讨

介绍了Cibacon FN染料的结构及染色性能,重点探讨了CibacronFN染料染大豆纤维/竹纤维混纺织物的条件,即染色温度为70℃,保温时间为60 min,染浅色时Na2SO4的用量为8～15 g/L,染中色时为15～15 g/L,染浅色时Na2SO3的用量为5～10 g/L,染中色时为13～15 g/L.确定了CibacronFN2R黄、CibacronFNR红、CibacronFNR蓝做三元色,对染色中存在的问题进行了分析,找出了原因,给出了解决办法.

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。

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.

天花粉蛋白与CibacronBlueF_3GA结合特性的研究

天花粉蛋白与ＣｉｂａｃｒｏｎＢｌｕｅＦ_３ＧＡ结合特性的研究何贤辉，柯一保，孙汛，聂慧玲（中国科学院上海细胞生物学研究所，上海２０００３１）天花粉蛋白（Ｔｒｉｃｈｏｓａｎｔｈｉｎ，简称ＴＣ８）是从葫芦科植物栝楼（Ｔｂｅｈｏｓａｎｔｈｅｓｋｉｒｉｌｏｗ?..

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.

Cibacron HW型活性深蓝代替硫化深蓝灯芯绒染色的实践

阐述了用ＣｉｂａｃｒｏｎＨＷ活性深蓝代替硫化深蓝的可行性及实际意义，并讨论了ＨＷ型活性深蓝染色的工艺要点。

CIBACRONS型染料在Modal/棉弹力针织物上的应用

文章介绍了CIBACRONS型染料的染色性能,重点探讨了CIBACRONS型染料在Modal/棉弹力针织物上染深浓色的工艺条件;分析了染色中存在的问题及原因,提出了解决办法。

Concentration and Preservation of Very Low Abundance Biomarkers in Urine, such as Human Growth Hormone (hGH), by Cibacron Blue F3G-A Loaded Hydrogel Particles

Urine is a potential source of diagnostic biomarkers for detection of diseases,and is a very attractive means of non-invasive biospecimen collection.Nonetheless,proteomic measurement in urine is very challenging because diagnostic biomarkers exist in very low concentration(usually below the sensitivity of common immunoassays)and may be subject to rapid degradation.Hydrogel nanoparticles functionalized with Cibacron Blue F3G-A(CB)have been applied to address these challenges for urine biomarker measurement.We chose one of the most difficult low abundance,but medically relevant,hormones in the urine:human growth hormone(hGH).The normal range of hGH in serum is 1 to 10 ng/mL but the urine concentration is suspected to be a thousand times less,well below the detection limit(50 pg/mL)of sensitive clinical hGH immunoassays.We demonstrate that CB particles can capture,preserve and concentrate hGH in urine at physiological salt and urea concentrations,so that hGH can be measured in the linear range of a clinical immunometric assay.Recombinant and cadaveric hGH were captured from synthetic and human urine,concentrated and measured with an Immulite chemiluminescent immunoassay.Values of hGH less than 0.05 ng/mL(the Immulite detection limit)were concentrated to 2 ng/mL,with a urine volume of 1 mL.Dose response studies using 10 mL of urine demonstrated that the concentration of hGH in the particle eluate was linearly dependent on the concentration of hGH in the starting solution,and that all hGH was removed from solution.Thus if the starting urine volume is 100 mL,the detection limit will be 0.1 pg/mL.Urine from a healthy donor whose serum hGH concentration was 1.34 ng/mL was studied in order detect endogenous hGH.Starting from a volume of 33 mL,the particle eluate had an hGH concentration of 58 pg/mL,giving an estimated initial concentration of hGH in urine of 0.175 pg/mL.The nanotechnology described here appears to have the desired precision,accuracy and sensitivity to support large scale clinical studies of urine hGH levels.

染料壳聚糖微球的制备及其对牛血清白蛋白(BSA)吸附性能的研究

采用反相悬浮交联法制备壳聚糖微球,对微球进行羟丙基氯化及氨基化,并偶联色素配体C ibacronB lue F3GA,得到一种新型染料亲和吸附剂.以牛血清白蛋白(BSA)为目标蛋白,考察了该染料亲和吸附剂的吸附性能,发现其对BSA有较高的吸附量(95.2 mg/g),吸附行为满足Langmu ir吸附等温式.负载牛血清白蛋白的微球容易洗脱,洗脱率高达99%.

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.

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 .

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.

用三嗪染料配基吐温80-磷酸盐液固萃取-体系分离纯化乳酸脱氢酶

目的 利用三嗪类染料 Cibacron Blue F3 G-A修饰的吐温 80 ,与磷酸钾盐构建染料配基吐温 80 -盐液 -固亲和萃取体系 ,从猪心肌匀浆液中分离纯化乳酸脱氢酶。方法 成相染料配基吐温 80浓度为 1 5 % (其中吐温 80中含染料配基吐温 80为 1 0 % ) ;成相磷酸钾盐浓度为 1 .49mol/L (p H 8.5 0 ) ;匀浆液加入量为体系的1 % ,一次成相正萃取。将吐温 80相保温溶解 ,改变磷酸钾盐浓度为 0 .91 mol/L,二次成相反萃取。结果 一次成相正萃取吐温 80相平均酶活收得率 97% ,纯化 8.0倍。二次成相反萃取 ,盐水相酶活收得率近 1 0 0 % ,实现了酶与吐温 80相的分离。结论 三嗪染料配基吐温 80 -磷酸盐液 -固萃取体系 ,分离纯化乳酸脱氢酶 ,具有良好选择性和分离纯化效果。对其进一步研究与开发 ,具有较大实际应用价值。