Q Sepharose^TM XL强阴离子交换色谱快速纯化猪心中肌红蛋白和细胞色素C

以Q SepharoseTM XL强阴离子交换色谱结合Sephadex G-75分子排阻色谱快速提取纯化猪心中肌红蛋白（Myoglobin,Mb）和细胞色素C（Cyt C）。在pH值为7.6-8.5范围内,Cyt C吸附在Q SepharoseTMXL强阴离子交换柱上,而Mb会缓慢通过。所以选择QSepharoseTMXL强阴离子交换柱的上样pH值为7.9,洗脱pH值为7.2。电泳检测证明,经过QSepharoseTMXL柱初步纯化所得Mb和Cyt C的产率分别为89.7%和93.2%;再经过Sephadex G-75分子排阻色谱可得到纯的Mb和Cyt C,其产量分别为77.43mg.（kg猪心）-1和86.42mg.（kg猪心）-1。使用QSepha-roseTMXL强阴离子交换色谱能从猪心中快速制备纯的Mb和Cyt C。     

Q Sepharose^TM-XL强阴离子交换色谱快速纯化精氨酸激酶及其多克隆抗体的研究

利用QSepharose^TM-XL强阴离子交换色谱从虾的精氨酸激酶粗提液中纯化出精氨酸激酶。以其免疫大白兔．然后利用QSepharose^TM-XL强阴离子交换色谱从高免疫兔血清中纯化精氨酸激酶的多克隆抗体。采用SDS-PAGE检测精氨酸激酶及其抗体的纯度,并进行了活性测定。结果表明．利用QSepharose^TM-XL强阴离子交换色谱纯化出的精氨酸激酶及其多克隆抗体纯度高、活性强。建立了快速高效、操作方便而又适合实际应用的制备精氨酸激酶及其多克隆抗体的方法．可为精氨酸激酶及食品过敏的相关研究提供帮助。     

以双水相萃取法从猪心中快速制备细胞色素C

把化工上常用的双水相萃取法应用于生物分离,从猪心中纯化具有药用价值的细胞色素C。以14％聚乙二醇和12％硫酸铵两相系统对粗提液进行萃取,细胞色素C全部浓缩于盐相。对盐相进行去盐后,经人造沸石快速吸附与洗脱、三氯醋酸沉淀等制得细胞色素C半成品,以弱酸性阳离子交换柱进行精制。聚丙烯酰胺凝胶电泳、可见吸收光谱扫描等定性定量分析证明了此法所得产品纯度好,提取率也较高。与传统方法比较,此法工艺简单,成本低,耗时少,尤其适用于大规模生产。     

全原子分子动力学模拟反相色谱分离细胞色素C的过程

采用全原子分子动力学方法模拟了反相色谱分离蛋白质的吸附和洗脱过程。采用表面键合C4烷基链的硅胶基质作为反相色谱介质和细胞色素C作为蛋白质模型,模拟蛋白质在反相色谱分离过程中的构象变化。结果显示:在吸附过程中,蛋白质在释放出表面结合水分子的同时置换出色谱介质表面的水分子。与其在溶液中的天然构象相比,吸附态蛋白质的构象发生改变。在洗脱过程中,随着溶剂从水切换到甲醇,甲醇取代水分子包覆在介质和蛋白质表面,将蛋白质从介质表面置换下来。分子模拟的结果再现了有关反相色谱\"优先水化\"的吸附机制,并从分子水平上展现了吸附和洗脱过程中蛋白质、色谱介质和溶剂之间相互作用,对反相色谱介质设计和过程优化提供了参考。     

高效阴离子交换色谱-脉冲安培法测定W135群和Y群脑膜炎球菌多糖中半乳糖和葡萄糖含量

目的采用高效阴离子交换色谱-脉冲安培法(High-performance anion-exchange chromatography with pulsedamperometric detection,HPAEC-PAD)测定W135群和Y群脑膜炎球菌荚膜多糖中半乳糖和葡萄糖的含量,并对该方法进行验证及初步应用。方法多糖蛋白结合物原液的混合液经HCl水解处理后,进样CarboPac PA10柱,以氢氧化钠-醋酸钠梯度流洗,脉冲积分安培检测器检测。筛选样品的水解条件;对HPAEC-PAD法进行专属性、精密性及准确性验证,并进行初步应用。结果确定的样品水解条件为以2 mol/L HCl于85℃处理1 h。载体蛋白破伤风类毒素对检测结果无干扰;重复性、日内精密性和不同操作者之间精密性试验结果的RSD均小于5%;准确性试验中葡萄糖和半乳糖的回收率在90%～110%之间。HPAEC-PAD法测定多糖蛋白结合物原液的多糖含量与传统间苯二酚比色法测定结果相当;成品疫苗中的W135群和Y群多糖含量测定结果符合《A、C、W135、Y群脑膜炎球菌结合疫苗制造及检定规程草案》的规定(均≥4μg/剂)。结论 HPAEC-PAD法专属性、精密性和准确性良好,可用于A、C、W135、Y群脑膜炎球菌结合疫苗中W135群和Y群多糖含量的测定。     

串联阴阳离子交换色谱分离牛初乳中乳铁蛋白与IgG

脱脂牛初乳用浓度为6 mol/L的盐酸调节其pH至4.0,以5 000 r/m in离心分离20 m in除去酪蛋白制得乳清,缓慢滴加浓度为1 mol/L的氢氧化钠回调pH至6.8;处理后的乳清经截留相对分子质量为50 000的组分并超滤稀释后,再经过阴阳离子交换串联色谱,乳铁蛋白(Lf)吸附于CM Sepharose Fast F low,免疫球蛋白G(Immunoglobu lin G,IgG)吸附于DEAE Sepharose Fast F low;阳离子交换柱用c(NaC l)=0.27 mol/L、0.85 mol/L的水溶液阶跃洗脱,得到色谱纯度为96.6%的Lf产品,阴离子交换柱用c(NaC l)=17 mmol/L、51 mmol/L的水溶液阶跃洗脱,得到色谱纯度为95%的IgG产品。     

Separation and Purification of Sinigrin and Gluconapin from Defatted Indian Mustard Seed Meals by Macroporous Anion Exchange Resin and Medium Pressure Liquid Chromatography

Defatted seed meals, the byproducts of Indian mustard seed oil industry, which are just used as animal feedstock and nitrogen fertilizers, possess various nutrients and phytochemical compounds, such as sinigrin and gluconapin. The present study demonstrated a novel, low-cost, and efficient method for the co-production of sinigrin and gluconapin from defatted Indian mustard seed meals by D261 strong basic anion-exchange macroporous resin and medium pressure liquid chromatography (MPLC). The adsorption/desorption capacities of eight adsorbents characterized by BET, IR, and EDS were screened. 26.4 g defatted Indian mustard seed meals, which contained about 1210.3 mg sinigrin and 696.6 mg gluconapin, could produce 2059.6 mg of glucosinolate-rich extract of 53.67% sinigrin and 31.30% gluconapin after separation by D261 resin. Then, 949.40 mg of 97% sinigrin and 568.40 mg of 95% gluconapin could be obtained after the extracts were purified by MPLC, with the recovery of 76.03% of sinigrin and 77.38% of gluconapin. The products were assessed by analytical HPLC and characterized by UV, MS, ¹H NMR, and ¹³C NMR. In conclusion, this method is practical and environmentally friendly, and it is a low-cost process to make full use of Indian mustard seeds.     

Mixed-Beds of Strong and Weak Anion Exchange Resins for Protein Separations with Step-Induced pH Gradients

A new way of performing pH gradient-based protein separations using a mixed-bed column comprising a small-pore weak base resin and a large-pore strong base resin and a step change in pH at the column entrance is introduced. The approach effectively decouples intracolumn generation of pH gradients, which ensues from ionic interactions with the weak base resin, from protein separation, which ensues from interactions with the strong base resin ligands, resulting in a process that is more easily predictable, less prone to the effects of protein loading on the pH gradient, less susceptible to hydrophobic interactions between protein and resin surface, and more flexible. The approach is demonstrated successfully using a tertiary amine resin as the weak base component together with a range of commercial strong anion exchangers with quaternary amine functionality. A predictive model, based on the potentiometric titration of the weak base resin, is in excellent agreement with experimentally determined pH profiles obtained using mixed-bed columns and amine buffers with pH steps from 9 to 7. The separation of protein charge variants with the mixed-bed concept is demonstrated using these columns for a deamidated monoclonal antibody mixture, which yielded highly homogeneous fractions.     

阴离子交换高效液相色谱法同时分离和分析三种氨基羧酸

建立了利用阴离子交换高效液相色谱同时分离分析甘氨酸、亚氨基二乙酸和氨三乙酸的方法。色谱柱为Sax强阴离子交换柱,用0.01 mol/L磷酸二氢钾缓冲液(pH=2.2)为流动相,紫外检测器波长为210 nm,对样品进行分析,并用外标法对这三种氨基羧酸进行定量。结果表明甘氨酸、亚氨基二乙酸和氨三乙酸分别在0.1024~2.0484 g/L、0.1006~2.0119 g/L、0.0050~0.3009g/L范围内线性关系良好(r>0.9995)。此法具有灵敏度高,线性相关系数和重现性好,流动相成本较低等特点。     

从阴树脂氯化母液中回收氧化锌技术

介绍了氯化母液中回收制取氧化锌的具体办法。     

Predicting Retention and Resolution of Protein Charge Variants in Mixed-Beds of Strong and Weak Anion Exchange Resins with Step-Induced pH Gradients

A model is developed to predict protein retention and resolution in mixed-bed columns containing a mixture of a small-pore weak base resin and a large-pore strong base resin. Using amine buffers, the weak base resin generates smooth pH gradients when the column is subjected to pH steps, while the strong base resin interacts with the proteins effecting the separation. The model describes simultaneously ion exchange interaction on the weak base resin and protein adsorption and transport on the strong base resin. The separation of a monoclonal antibody (mAb) from a charge variant whose pI is about 0.2 pH units lower is studied experimentally with the anion exchange resins AG 4-X4 and SOURCE 30Q as the weak and strong base components, respectively. The properties of each resin are characterized independently and model parameters are obtained experimentally for each. Model predictions of pH gradient-based separations in columns containing different proportions of the two resins and at different buffer concentrations are found to be in agreement with experimental results. The model can thus be used as a process design tool to predict combinations of buffer and mixed-bed compositions, column lengths, and mobile phase velocities that would yield a desired separation performance.     

改性季铵化壳聚糖阴离子交换膜的制备与表征

以壳聚糖（CS）为原料,2,3-环氧丙基三甲基氯化铵（GTA）为醚化剂,通过亲核取代反应制备了季铵化壳聚糖（QCS）,用傅立叶变换红外光谱（FT-IR）表征了产物的结构,结果表明产物结构与目标产物的结构相符。以QCS为原料,醋酸溶液为溶剂,通过加入交联剂戊二醛（GA）和荷正电聚苯乙烯（PS）微球,制备了一系列改性QCS阴离子交换膜,并对其含水率、溶胀度、离子交换量等性能进行了测定。结果表明：交联剂的加入可有效抑制QCS膜的形变,降低了其溶胀度,微乳液的加入会在一定程度上提高复合膜的离子交换能力,交联度为4%、微乳液用量为15%（v/v）时离子交换膜的含水率趋于稳定。     

Selective Separation and Recovery of Zinc and Lead from Galvanizing Industrial Effluents by Anion Exchange

The recovery of zinc and its separation from iron, hydrochloric acid, and minor elements in the galvanizing pickling baths were studied by anion exchange. The breakthrough curves demonstrated that the resins, Lewatit MonoPlus MP 500 and AMBERLYST A21, exhibit a greater capacity for zinc than for cadmium, copper, and lead. The results indicated the feasibility of operating the ion exchange process to treat a real effluent. During the loading, lead started eluting until its complete removal from the resin. Like this, a two-step removal solution is proposed, using two columns with Lewatit MonoPlus MP 500, for separating zinc and lead.     

Production of Neptunium Oxide from Impure Solutions by Anion Exchange Followed by Oxalate Precipitation

Abstract

The stabilization of the neptunium solutions stored at the Savannah River Site (SRS) has generated additional recycle solutions that contain a different mix of impurities. A list of expected purities and the known laboratory and production data for purification from those impurities has been accumulated. An evaluation has been performed of the options for modifying the current process to ensure oxide product purity does not significantly change when recycle solutions are processed. This paper discusses the details of the reduction of the major impurities utilizing both known production quality analyses and laboratory flowsheet development data and proposes modifications to the anion wash volumes to remove higher levels of each impurity.