反离子对部分电荷中和的聚乙烯亚胺接枝介质的蛋白质色谱行为的影响

前期研究表明部分电荷中和的聚乙烯亚胺接枝琼脂糖介质FF-PEI-R440具有较高静态吸附容量和快速传质速率。本文选取硫氰酸根离子、氯离子、磷酸氢根离子以及硫酸根离子这4种反离子,在离子强度为0.03、0.06 mol·L^-1下,研究其对FF-PEI-R440吸附与洗脱行为的影响。结果表明：在两种离子强度下,饱和吸附容量的增加顺序均为SCN^- < SO₄^2- < HPO₄^2- < Cl^-;在低离子强度下,除SCN^-外,反离子种类对传质速率和动态结合容量没有显著影响;在高离子强度下,传质速率的增加顺序为SCN^- < SO₄^2- ≈ Cl^- < HPO₄^2-,而动态结合容量的增加顺序与饱和吸附容量序列一致;两种离子强度下Cl^-均保持最高的动态结合容量;反离子种类对蛋白质洗脱行为没有显著影响。上述结果说明反离子主要影响FF-PEI-R440的吸附性能,而不影响洗脱,且流动相中选择Cl^-为反离子最利于FF-PEI-R440的实际柱色谱操作。     

接枝聚合物配基的蛋白质吸附层析

以离子交换、亲和结合和疏水性吸附为主的蛋白质吸附层析是药用蛋白质生产过程的核心技术,开发新技术和提高蛋白质吸附层析操作的分离效率(如选择性和动态吸附容量等)是该领域的主要研究目标。近年来聚合物配基接枝的层析介质由于同时具有较高的吸附容量和传质速率,得到产学界的广泛关注。本文针对聚合物配基接枝修饰的蛋白质吸附层析介质的配基化学特征、吸附和传质特性、层析分离应用和设计等方面进行评述。首先介绍不同种类的聚合物接枝介质,然后系统阐述聚合物配基化学特性对介质吸附和传质性能的影响机制,并分析上述性质对聚合物配基接枝层析介质分离特性的影响机理和应用,最后讨论和展望了高效聚合物配基接枝介质设计、开发和应用的前景。     

葡聚糖接枝型耐碱Protein A介质的层析性能提升研究

Sepharose 4FF微球经环氧活化后与葡聚糖溶液反应,得葡聚糖接枝型琼脂糖微球,再经环氧活化和偶联耐碱型Protein A配基,得葡聚糖接枝型高载量Protein A介质,测定了介质在线清洗稳定性能,并进行了热力学研究. 结果表明,与常规Protein A介质相比,葡聚糖接枝型Protein A介质的最高流速提高约32%,对抗体hIgG的动态载量为60.6 mg/mL,分别为常规介质和MabSelect SuRe介质载量的123%和95%;经40次清洗后,葡聚糖接枝型Protein A介质动态载量为原始载量的92%,远高于常规介质的84%,与MabSelect SuRe稳定性基本一致. 3种介质对抗体的结合均为熵驱动过程,葡聚糖接枝型Protein A介质的吸附热介于MabSelect SuRe和常规Protein A介质之间.     

反离子对部分电荷中和的聚乙烯亚胺接枝介质的蛋白质色谱行为的影响

前期研究表明部分电荷中和的聚乙烯亚胺接枝琼脂糖介质FF-PEI-R440具有较高静态吸附容量和快速传质速率。本文选取硫氰酸根离子、氯离子、磷酸氢根离子以及硫酸根离子这4种反离子,在离子强度为0.03、0.06mol·L~(-1)下,研究其对FF-PEI-R440吸附与洗脱行为的影响。结果表明:在两种离子强度下,饱和吸附容量的增加顺序均为SCN-SO_4~(2-)HPO42-Cl~-;在低离子强度下,除SCN-外,反离子种类对传质速率和动态结合容量没有显著影响;在高离子强度下,传质速率的增加顺序为SCN-SO_4~(2-)≈Cl~-HPO42-,而动态结合容量的增加顺序与饱和吸附容量序列一致;两种离子强度下Cl~-均保持最高的动态结合容量;反离子种类对蛋白质洗脱行为没有显著影响。上述结果说明反离子主要影响FF-PEI-R440的吸附性能,而不影响洗脱,且流动相中选择Cl~-为反离子最利于FF-PEI-R440的实际柱色谱操作。     

葡聚糖接枝型Protein A介质的层析性能提升

Sepharose 4FF微球经环氧活化后与葡聚糖溶液反应,得葡聚糖接枝型琼脂糖微球,再经环氧活化和偶联耐碱型Protein A配基,得葡聚糖接枝型高载量Protein A介质,测定了介质在线清洗稳定性能,并进行了热力学研究.结果表明,与常规Protein A介质相比,葡聚糖接枝型Protein A介质的最高流速提高约32%,对抗体h Ig G的动态载量为60.6 mg/m L,分别为常规介质和Mab Select Su Re介质载量的123%和95%;经40次清洗后,葡聚糖接枝型Protein A介质动态载量为原始载量的92%,远高于常规介质的84%,与Mab Select Su Re稳定性基本一致.3种介质对抗体的结合均为熵驱动过程,葡聚糖接枝型Protein A介质的吸附热介于Mab Select Su Re和常规Protein A介质之间.     

改性MCM-41分子筛对阴离子偶氮染料的吸附作用机理

制备了改性介孔分子筛NH₃⁺-MCM-41,研究了其对阴离子偶氮染料甲基红的吸附效果。研究表明,通过氨基改性使介孔分子筛MCM-41表面电荷发生改变,从而利用静电吸附作用改善了脱除效果,使甲基红吸附率提高了1.1倍,达到90.4%。溶液pH值的改变会影响NH₃⁺-MCM-41的表面电荷,从而影响吸附效率,其最佳pH值为4。溶液中的杂质离子均会对吸附产生影响,当溶液中含有Cl^-、NO₃^-时,对吸附效果影响不大;而当溶液中含有CO₃^2-和HPO₄^2-等高价弱酸根离子时,对吸附效果影响很大,基本不吸附染料。此外,通过其吸附动力学研究发现,吸附过程中不符合准一级动力学模型,而较好符合准二级动力学,说明在该过程中存在化学吸附,这种化学吸附涉及吸附剂与吸附质之间的电子共用或电子转移。     

改进葡聚糖接枝技术制备高性能层析介质

利用改进的葡聚糖接枝技术,在以环氧氯丙烷为交联剂交联琼脂糖微球骨架的过程中加入葡聚糖溶液,在交联的同时接枝葡聚糖制得葡聚糖接枝型琼脂糖微球Rigose-Dex,再与盐酸2-氯三乙胺（DEAE）反应,获得葡聚糖接枝型高载量弱阴离子交换介质Rigose-Dex DEAE。以牛血清白蛋白（BSA）为模型蛋白,以商品化介质DEAE Sepharose 6FF为对照,系统研究了该葡聚糖接枝型Rigose-Dex DEAE的蛋白吸附性能,并进行了物理性能研究。结果表明,改进后葡聚糖接枝技术的最高接枝量为24.5mg/mL。自制Rigose-Dex DEAE可耐受700cm/h 的线性流速,对BSA的动态饱和载量为127.6mg/mL, 为商品介质DEAE Sepharose 6FF 载量的212%;具有在高流速下快速结合蛋白的能力,上样蛋白溶液在层析柱中停留2min即可基本达到饱和动态载量;重复使用性能好,经120 次在线清洗后,Rigose-Dex DEAE介质的动态载量为原始载量的90.4%。     

减电荷聚甲基丙烯酸钠接枝介质的制备及蛋白质吸附性能

离子交换容量（IC）为320mmol/L的聚甲基丙烯酸钠（pMA）接枝型介质（FF-pMA-320）对溶菌酶和γ-球蛋白具有较高的吸附容量，但其传质速率较低。在保持聚合物链长度的前提下，通过乙醇胺与接枝链上的羧基进行电荷中和反应，降低pMA接枝型介质的电荷密度，提升介质的蛋白质传质速率。将FF-pMA-320进行部分电荷中和修饰，制备得到离子交换容量分别为230mmol/L和170mmol/L的减电荷阳离子交换介质，分别命名为pMA-320-R230和pMA-320-R170。采用吸附平衡、吸附动力学和柱穿透实验，研究了溶菌酶和γ-球蛋白在这两种新型介质上的吸附行为，并与初始介质FF-pMA-320进行了比较。结果表明：随着介质的IC值（电荷密度）从320mmol/L降低到170mmol/L，介质对两种蛋白质吸附容量随之减少，这与电荷中和修饰降低蛋白质吸附位点有关。随着接枝聚合物电荷密度的降低，相邻聚合物链之间的静电排斥作用减弱，蛋白质吸附容量降低，造成接枝链的灵活性增加以及蛋白质排阻效应减弱。因此，溶菌酶和γ-球蛋白在pMA-320-R170上的传质速率分别是FF-pMA-320的1.6倍和5.5倍。柱穿透实验结果表明，由于pMA-320-R170对γ-球蛋白传质速率较高，使得介质对蛋白质的动态结合容量（DBC）高于其他两种介质，在流速为150~750cm/h时可保持在10mg/mL以上。研究结果对设计和开发高性能蛋白质色谱介质提供借鉴和指导。     

焦化脱硫废杂盐资源化新技术

由于HPF法焦炉煤气脱硫工艺会产生危害极大的脱硫废杂盐,本文以回避传统提盐法存在的工艺流程长、产品收率低等难题为出发点,提出硫酸铜沉淀硫氰酸根、微纳米气泡氧化硫代硫酸根、石灰表面强制沉淀硫酸根同步制氨的新技术。在实验室配制模拟脱硫废杂盐,优化新技术反应条件,实验表明,当[Cu²⁺]∶[SCN^-]摩尔比为1.2、温度为40℃、初始废液硫氰酸根浓度大于300g/L、反应80min后,SCN^-最佳去除率为99.20%;当pH=1、温度为50℃、初始废液硫代硫酸根浓度为50g/L、反应420min后,S₂O₃^2-去除率为95.18%;当[Ca²⁺]∶[SO₄^2-]摩尔比为1.5、反应温度为20℃、初始废液硫酸根浓度为500g/L,并加入5g直径5mm PP球充当研磨介质,反应240min后,SO₄^2-去除率为91.11%。     

葡聚糖接枝型高载量金属螯合介质的制备与性能

对琼脂糖凝胶微球进行烯丙基活化,再接枝葡聚糖分子,考察葡聚糖分子量等因素对葡聚糖接枝过程的影响;以葡聚糖接枝琼脂糖凝胶微球为基质,制备亚氨基二乙酸型金属螯合介质,考察葡聚糖接枝过程对金属螯合介质的孔道结构、流通性能和载量等的影响. 结果表明,分子量20~500 kDa的葡聚糖都能均匀分布于琼脂糖凝胶微球内,葡聚糖接枝量随分子量增加而增大,所制的金属螯合介质形貌、粒径及其分布基本不受影响,且具有更好的流通性能,孔道结构比商品介质Ni Sepharose 6FF更丰富. 葡聚糖接枝的金属螯合介质对带组氨酸标签的乳酸脱氢酶和睫状神经营养因子的载量分别达到19和27 mg/mL,较Ni Sepharose 6FF的载量分别提高26.6%和42.0%.     

Protein adsorption onto diethylaminoethyl dextran modified anion exchanger: Effect of ionic strength and column behavior

Our previous studies on bovine serum albumin(BSA) adsorption to diethylaminoethyl dextran(DEAE dextran,DexD, grafting-ligand) and DEAE(D, surface-ligand) modified Sepharose FF resins found that all the grafted resins(FF-DexD and FF-D-DexD) exhibited extremely fast uptake rate(effective diffusivity, D_e, D_e/D_O 1.4),which was six times greater than the ungrafted resins(D_e/D_O 0.3). In this work, the influence of ionic strength(IS) on 6 typical DEAE dextran-grafted resins was investigated. Bath adsorption equilibria and kinetics, breakthrough, and linear gradient elution experiments were conducted. Commercial DEAE Sepharose FF was used for comparison. It is found that protein adsorption capacities on DEAE dextran-FF resins and the commercial resin decreased with increasing IS, but DEAE dextran-FF resins exhibited much higher capacity sensitivity to salt concentration. Besides, steeper decrease of adsorption capacities could be obtained at higher graftingligand or surface-ligand density. It is worth noting that the facilitating role of surface-ligand to the "chain delivery" effect was weakened after adding salt, leading to the less improvement in uptake rate by increasing surface-ligand density at higher IS. Although the uptake rates of the DEAE dextran-FF resins increased first and then decreased with increasing IS, they kept the extremely high level of De values(D_e/D_O 1.1) at the their working/binding IS range. Moreover, the DEAE dextran-FF resin displayed much higher adsorption capacities and De values than commercial ungrafted resin in their working condition. Furthermore, the column results of DEAE dextran-FF resins presented higher dynamic binding capacities than and similar elution ISs with DEAE Sepharose FF to achieve similar(or even higher) recoveries suggest the excellent chromatographic column performance of the DEAE dextran-FF resins. Finally, both high recovery and purity of BSA and γ-globulin could be easily achieved using the typical DEAE dextran-FF column, FF-D60-DexD160, to separate their binary mixtures,by step gradient elution. The research has provided new insights into the practical application of the series of DEAE-dextran grafted resins in protein chromatography and proved their superiority.     

Protein retention in dextran-grafted cation exchange chromatography: The influence of pHs,counterions and polymer structure

Polymer-grafted ion exchange adsorbents were of great interest for the development of high-performance protein chromatography in biopharmaceutical and related fields. In this work, protein retention was systematically investigated in ion exchange chromatography packed respectively with dextran-grafted cation exchange adsorbents containing sulphopropyl(SP) ligand, SP Sepharose XL and Capto S, and non-grafted cation exchange adsorbent, SP Sepharose FF, using five proteins. With an increase of buffer p Hs, retention factors of proteins decreased among all the adsorbents, demonstrating the dominant role of electrostatic interaction for protein binding on cation exchange adsorbents. The evidences further revealed that the scattered positive charges on the surface of protein molecules, rather than net charge of protein molecule, determined protein retention on cation exchange adsorbent. Likely, counterions including NH~(4+), K~+, Na~+ and Mg~(2+) exhibited distinct influence on protein retention. It was well ascribed to solvent-mediated indirect ion–macromolecule interactions and direct ion–macromolecule interactions. Compared with SP Sepharose FF, polymer structure in dextran-grafted cation exchange adsorbents ultimately brought about different ligand distributions and smaller pore sizes, thereby regulating protein retention in cation exchange chromatography. By comparing the retention of myoglobin and β-lactoglobulin B in SP Sepharose XL and Capto S, we reasonably speculated that the enhancement of nonelectrostatic interaction caused by reducing the space arm length was a major reason for an increasing retention factor of myoglobin in Capto S. The results in this research help us understand adsorption mechanism of protein in polymer-grafted adsorbents and give scientific guidance for the development of chromatographic materials.     

A222型阴离子交换树脂脱除HPPH+体系中的SO42-、NO3-

优化选取凝胶型强碱性苯乙烯系A222型阴离子交换树脂,对HPPH⁺溶液中SO₄^2-、NO₃^-的交换性能进行了研究。通过静态法分别考察了A222型阴离子交换树脂在水溶液及0.05mol/L HPPH⁺溶液中对SO₄^2-和NO^3-脱除过程。结果表明,A222型阴离子交换树脂对HPPH⁺无化学吸附作用,在水体系中对SO₄^2-和NO₃^-的平衡交换量分别为0.8382mmol/g和1.2980mmol/g,符合Langmuir和Freundlich吸附经验方程。树脂用量和温度是离子交换过程的主要影响因素,温度升高有利于离子交换的进行,树脂的优化用量为4g/L。A222型阴离子交换树脂可高效联合脱除HPPH⁺体系中的SO₄^2-和NO₃^-。FTIR谱图表征显示,SO₄^2-和NO₃^-可与树脂交换基团进行交换。     

Implications from γ-globulin adsorption onto cation exchangers fabricated by sequential alginate grafting and sulfonation

Our previous work proved that high adsorption capacity and uptake rate of lysozyme were achieved on alginate(Alg)-grafted re sin with an ionic capacity(IC) of 240 mmol·L~(-1)(Alg-FF-240).Moreover,the salt-tolerant feature of Alg-FF-230 was improved by using sequential alginate grafting and sulfonation strategy.Inspired by the enhanced adsorption performance of lysozyme,we have herein proposed to investigate the static and dynamic adsorption behaviors of γ-globulin on a series of Alg-grafted resins with different grafting densities and sulfonation degrees.The adsorption ca pacity of γ-globulin decreased with increa sing alginate-grafting density(IC) from 160 to 230 mmol·L~(-1) at 0 mmol·L~(-1) NaCl because of the steric hindrance caused by the alginate-grafting layer.Effects of ionic strength(IS) indicated that the adsorption capacities of the resins with the IC value of 230-370 mmol·L~(-1) were much higher than CM Sepharose FF at 50-100 mmol·L~(-1) NaCl,and the uptake rate of Alg-FF-230 was about twice as much as that of CM Sepharose FF.This work demonstrated the important effects of alginate-grafting layer and IS in γ-globulin adsorption behavior,which would be helpful in the design of Alggrafted resins and the selection of proper IS condition for protein purification.     

CuSO4对氰化提金贫液中SCN-的沉淀效果以及对共存离子浓度的影响

针对氰化提金贫液循环利用过程SCN^-积累以及外排带来的环境污染问题，以CuSO₄作为沉淀剂处理某高浓度SCN^-贫液，考察了沉淀剂用量、沉淀时间及沉淀温度对SCN^-沉淀效果和对共存离子浓度的影响。结果表明，当在贫液中加入1.6倍化学反应计量的CuSO₄后于室温搅拌60 min，SCN^-的去除率达87.5%以上，体系中T_Cu、S₂O₃ ^2-和SO₃ ^2-浓度也大幅降低，同时得到纯度达97%以上的CuSCN产品。沉SCN^-后的贫液只需石灰中和并沉部分SO₄ ^2-后，其活性得到进一步恢复，可直接返回金精矿浸出工序充分利用其中的CN^-，实现提金贫液的循环利用。     

Preparation and characterization of sulfated TiO2 with rhodium modification used in esterification reaction and decomposition of methyl orange☆

A unique Rh/TiO_2 solid acid catalyst modified with H_2SO_4 was synthesized and evaluated in the esterification reaction of propylene glycol methyl ether and decomposition of methyl orange(MO) in aqueous phase under halogen lamp irradiation. For this purpose, rhodium(Rh) nanoparticles were loaded on SO_4~(2-)/TiO_2 via the photo-deposition method. It was found that SO_4~(2-)/Rh–TiO_2 exhibited stronger catalytic activity than SO_4~(2-)/TiO_2. The new catalysts were characterized by X-ray powder diffraction(XRD), Brunauer–Emmett–Teller(BET), Transmission electron microscopy(TEM) and high-resolution(HRTEM), X-ray photoelectron spectroscopy(XPS) and Fourier Transform infrared spectroscopy(FTIR). Results from XRD and BET show that SO_4~(2-)/Rh–TiO_2 has higher specific surface area and smaller pore size than SO42-/TiO_2. The distribution of loaded Rh was found to be uniform with a particle size of 2–4 nm. Data from XPS reveal that Rh primarily exists as Rh~0 and Rh~(3+)in Rh–TiO_2 and SO_4~(2)-/Rh–TiO_2. These valence forms of Rh likely contribute to the enhanced catalytic activity. Furthermore, FT-IR spectra of the catalysts show an abundance of surface hydroxyl groups, which help the formation of hydroxyl radicals and the enhancement of surface acid density. The results show that more acid sites are formed on the sulfated Rh–TiO_2, and these acidic sites are largely responsible for improving the catalytic performance. This superior SO_4~(2-)/Rh–TiO_2 catalyst has potential applications in reactions requiring efficient acid catalysts, including esterification reactions and waste water treatment.     

硫酸根对聚合氮化碳光催化活性的影响

在pH=1条件下分别加入硫酸根、硝酸根、磷酸根对三聚氰胺进行处理,采用高温煅烧法制备了一系列聚合氮化碳材料,并通过X射线衍射（XRD）、扫描电子显微镜（SEM）、紫外-可见漫反射光谱（UV-Vis DRS）等分析手段对样品进行了表征。以罗丹明B（RhB）为污染物,探究了在可见光下聚合氮化碳对RhB的光催化降解性能、稳定性以及降解机理。加入0.12 mol硫酸根处理的聚合氮化碳（CN-0.04SO₄^2-）在45 min内对RhB的降解率为99.1%,其伪一级动力学常数是未加硫酸根的23.0倍,硫酸根能有效地增强聚合氮化碳的光催化活性。CN-0.04SO₄^2-经过3次循环实验对RhB的降解率略有下降,说明其具有良好的稳定性。活性物种捕获实验表明,影响光催化降解RhB的主要活性物种是·O₂^-和·OH。