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1.
针对色谱分离过程优化,建立了基于微孔过滤板的蛋白吸附高通量筛选方法,用于介质初筛、吸附性能考察、吸附等温线和吸附动力学测定、吸附和洗脱条件优化等。首先优化了96孔过滤板的操作参数,以2种离子交换介质和2种混合模式介质为典型代表,采用微孔过滤板方法考察了不同介质和液相条件下牛血清白蛋白的吸附,得到结合载量分布图,确定了合适的蛋白吸附和解吸条件。进一步测定了4种介质在特定吸附条件下的吸附等温线和吸附动力学曲线,获得吸附相关参数。最后,采用微孔过滤板进行了洗脱条件优化,并与填充柱色谱分离进行比较,验证了方法的可靠性。结果表明,基于微孔过滤板的蛋白吸附高通量筛选是切实可行的,可以快速筛选色谱介质和液相,优化蛋白分离条件,具有资源消耗小、实验通量大、研发周期短、适用性广、稳定性高的特点,是蛋白色谱分离过程优化的一种新方法。  相似文献   

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用丙烯酰胺等离子体引发活化聚苯乙烯(PS)微孔板,接枝4-(N-马来酰亚胺基甲基)环己 烷-1-羧酸琥珀酰亚胺酯(SMCC)联接剂,制备了易于固定蛋白的PS-g-SMCC微孔板。利用FTIR、EDS、XPS、SEM、接触角测试仪和酶标仪对其表面组成、结构、形貌及蛋白吸附性能进行了表征。结果表明,PS微孔板表面成功接枝了丙烯酰胺与SMCC,当等离子体处理参数为500 W,300 s时,接枝丙烯酰胺的效果最好,PS-g-SMCC微孔板表面达到超亲水;当SMCC质量浓度为5 g/L 时,PS-g-SMCC微孔板对牛血清蛋白的吸附量最大,为903.08 ng/cm2,与未处理的PS微孔板相比,吸附量提高了2.93倍,有效提高了PS微孔板的检测灵敏性。  相似文献   

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本文考察了烧成温度、成形压力、烧成时间、成孔剂的加入量和玻璃相加入量对SiC微孔陶瓷过滤板孔隙率及微观结构的影响。研究表明:烧成温度为1300℃、成形压力为40kN、烧成时间为4h、成孔剂加入量为16.5%时,制备出的SiC微孔陶瓷过滤板有着更为优良的性能。  相似文献   

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本文考察了SiC微孔陶瓷过滤板的烧成温度、成型压力、烧成时间、成孔剂的加入量和玻璃相加入量对SiC微孔陶瓷过滤板孔隙率及微观结构的影响。研究表明:在烧成温度为1300℃、成型压力为40kN、烧成时间为4h,成孔剂和玻璃相加入量分别在16.5%、6%的条件下,制备出的SiC微孔陶瓷过滤板有着更为优良的性能。  相似文献   

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固液分离用微孔陶瓷过滤板的性能设计及制备   总被引:9,自引:0,他引:9  
简述了固液分离用陶瓷过滤板的高效节能机理 ,讨论了微孔陶瓷性能设计的关键要素 ,介绍了陶瓷过滤板的制备工艺。  相似文献   

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荷电微孔滤膜及其吸附性能的研究   总被引:7,自引:0,他引:7  
本文研究了以天然多糖为荷电剂,用浸涂法制备的荷电微孔滤膜的吸附性能,探讨了制膜工艺及操作条件,如溶液组分,温度及pH值等吸附性能的影响,为荷电微孔滤膜的使用提供依据。  相似文献   

8.
微孔陶瓷加压过滤的试验研究   总被引:1,自引:0,他引:1  
王甲  何振永 《过滤与分离》1996,(1):12-14,37
随着加压过滤机在我国的推广应用,过滤耗风显得尤为突出。本文论述了微孔陶瓷加压的过滤的原理、试验方法及实验室试验结果。  相似文献   

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微孔过滤介质的进展   总被引:2,自引:0,他引:2  
近20年来用于固液分离的普通过滤介质除用化纤织物几乎取代了天然织物过滤介质外,微孔过滤介质有了突破性进展,相继出现了亲水微孔陶瓷;微孔复合薄膜,以及单丝纤维过滤介质等,本文综合介绍它们的特性及其出现的背景和重要意义。  相似文献   

10.
康勇  魏巧莲  姚鹏  罗茜  胡筱敏 《化工进展》2004,23(4):412-417
对新型微孔薄膜复合过滤介质的过滤性能进行了系统研究,并与传统机织滤布的相关性能进行了对比。研究结果表明:新型微孔薄膜复合过滤介质具有极高的固体截留率、良好的抗堵塞性能和较长的使用寿命;在试验所考查的范围内,过滤介质的过滤速度稳定,基本不受过滤时间的影响;滤液浊度较低,基本不超过50mg/L;新型微孔薄膜复合过滤介质卸饼率高;气压过滤时的气耗量仅是普通机织滤布的0.056~0.11倍。  相似文献   

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A novel monolithic material was developed to obtain efficient and cost‐effective purification of IgG from human plasma. The porous monolith was obtained by bulk polymerization in a glass tube of 2‐hydroxyethyl methacrylate (HEMA) and N‐methacryloyl‐(L )‐histidine methyl ester (MAH). The poly(HEMA‐MAH) monolith had a specific surface area of 214.6 m2/g and was characterized by swelling studies, porosity measurement, FTIR, scanning electron microscopy, and elemental analysis. Then the monolith was loaded with Cu2+ ions to form the metal chelate. Poly(HEMA‐MAH) monolith with a swelling ratio of 74% and containing 20.9 μmol MAH/g was used in the adsorption/desorption of IgG from aqueous solutions and human plasma. The maximum adsorption of IgG from an aqueous solution in phosphate buffer was 10.8 mg/g at pH 7.0. Higher adsorption was obtained from human plasma (up to 104.2 mg/g), with a purity of 94.1%. It was observed that IgG could be repeatedly adsorbed and desorbed with the poly(HEMA‐MAH) monolith without significant loss of adsorption capacity. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 101: 395–404, 2006  相似文献   

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Penicillin G acylase (PGA) is one of the most important enzymes for the production of semi-synthetic β-lactam antibiotics and their key intermediates. Purification of penicillin G acylase from fermentation broth with the aid of high-throughput screening (HTS) process has been examined in this study. We used a microtiter-plate based on screening method to find appropriate purification conditions for the target protein. The screening method is based on a 96-well plate format, and different matrices and conditions (pH, salt concentration and type) were tested. Through analyses of all pooled fractions (flow-through and elution) we gained appropriate information to choose the best performing matrix and buffer conditions for upscaling. After an upscaled purification step the second unit operation is screened in the similar way and parameters for this operation can be chosen. The purification parameter of purified PGA at the small-screen and upscaling levels were measured, respectively. The results indicate that high-throughput progress based on a 96-well plate is a flexible and efficient paradigm for recombinant protein purification.  相似文献   

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存在于建筑材料中的氡(Rn)已成为污染室内空气的一种重要放射性气体,对人体的健康可造成严重的危害,因此开发对其具有高效分离性能的新型多孔材料具有重要的意义。基于巨正则系综的Monte Carlo模拟方法,系统地研究了163种锆基金属-有机骨架材料(Zr-MOF)在常温常压下对Rn/N2和Rn/O2混合气体的吸附分离性能。研究结果表明材料的孔径在5.6~8 ?(1 ? =0.1 nm)、可接触比表面积在140 ~870 m2/g范围内,材料对Rn的分离效果最佳,并发现在材料骨架上引入强极性功能基团如羧基(—COOH)和硫磺基(—SO3H)等,有利于强化材料对Rn的分离性能。研究结果可为今后理性设计与可控合成相关高性能MOF分离材料提供理论参考。  相似文献   

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己烷异构体中双支链异构体的分离可以提高汽油的辛烷值从而减少发动机的爆震现象。针对传统的蒸馏方法耗能高和新型吸附剂金属有机框架成本高、工作能力低、稳定性差的缺点,采用高通量计算筛选方法研究了688种共价有机框架(COFs)对己烷异构体的分离性能。首先计算了所有COF的几何结构描述符,通过限制孔径(PLD) 6.2~15?的范围筛选出209个可容纳所有己烷异构体的COF,再利用巨正则Monte Carlo (GCMC)方法模拟433 K下上述COF对己烷异构体的吸附解吸过程。对再生能力R>80%且吸附性能分值(APS)最高的COF进行排序,筛选出具有最高APS值的COF-DL229 2-fold,APS值为23.36 mol/kg,R为99.38%。分析了6个几何结构描述符与APS的相关性,发现对于COF来说较高的孔隙率(VF)、较高的孔隙体积(PV)、较低的密度(ρ)可提高COF的APS值。最后基于PV、VF、ρ利用决策树算法设计出高APS值COF的筛选路径,研究工作对今后设计用于己烷异构体吸附分离的COF具有一定的指导意义。  相似文献   

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Concanavalin A (Con A) immobilized poly(2‐hydroxyethyl methacrylate) (PHEMA) beads in a spherical form (100–150 μm in diameter) were used for the affinity chromatography purification of human immunoglobulin G (IgG) from aqueous solutions and human plasma. PHEMA adsorbents were prepared by suspension polymerization. Bioligand Con A was then immobilized by covalent binding onto PHEMA beads. The maximum IgG adsorption on the PHEMA/Con A beads was observed at pH 6.0. The nonspecific IgG adsorption onto the plain PHEMA adsorbents was very low (ca. 0.17 mg/g). Higher adsorption values (up to 54.3 mg/g) were obtained when the PHEMA/Con A beads were used from aqueous solutions. A higher adsorption capacity was observed for human plasma (up to 69.4 mg/g) with a purity of 82.5%. The adsorption capacities of other blood proteins were 2.0 mg/g for fibrinogen and 4.2 mg/g for albumin. The total protein adsorption was determined to be 76.0 mg/g. IgG molecules could be repeatedly adsorbed and desorbed with the PHEMA/Con A beads without noticeable loss in the IgG adsorption capacity. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 97: 1202–1208, 2005  相似文献   

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Most traditional D2/H2 separation techniques are energy-intensive with low efficiency. Metal–organic frameworks (MOFs) provide a promising solution for D2/H2 separation due to their excellent chemical and structural characteristics. Here, machine learning-assisted high-throughput computational screening was employed to identify the high-performance MOFs for the dynamic D2/H2 separation. Extensive data analysis reveals that there were two adsorption behaviors in the optimal MOFs, independent adsorption and competitive adsorption, and the independent adsorption was favorable for the preferential adsorption of D2. To quantify these two adsorption behaviors, we introduced and defined overlap degree (OD) and independence degree (ID), and developed a software for the rapid assessment of OD/ID. After batch simulation of the breakthrough curves of 2000 optimal MOFs, ~80% MOFs exhibited independent occupancy, confirming its contribution to good dynamic separation capabilities. This work provides a new idea for designing MOFs with independent adsorption behavior to improve the dynamic separation performance of hydrogen isotopes.  相似文献   

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Many protein–protein and peptide–protein interactions (PPIs) play key roles in the regulation of biological functions, and therefore, the modulation of PPIs has become an attractive target of new drug development. Although a number of PPIs have already been identified, over 100 000 unknown PPIs are predicted to exist. To uncover such unknown PPIs, it is important to devise a conceptually distinct method from that of currently available methods. Herein, an mRNA display by using a total RNA library derived from various human tissues, which serves as a unique method to physically isolate peptide epitopes that potentially bind to a target protein of interest, is reported. In this study, selection was performed against Kelch-like ECH-associated protein (Keap1) as a model target protein, leading to a peptide epitope originating from astrotactin-1 (ASTN1). It turned out that this ASTN1 peptide was able to interact with Keap1 more strongly than that with a known peptide derived from Nrf2; a well-known, naturally occurring Keap1 binder. This case study demonstrates the applicability of peptidomic mRNA display for the rapid exploration of consensus binding peptide motifs and the potential for the discovery of unknown PPIs with other proteins of interest.  相似文献   

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非平衡循环吸附色谱分离研究   总被引:3,自引:1,他引:2  
李忠  舒文利 《化工学报》1998,49(4):501-505
引言制备或大型液相色谱因在温和条件下具有高分辨分离能力且操作简单而广泛应用于药物、生物化学品和碳氢化合物等的分离和高纯化.其应用的范围和规模正在不断发展.然而,传统的吸附色谱因其吸附剂利用率低,生产效率低.为克服这一缺陷,Wankat,Barker和Ganetsos以及Ruthven和Ching等人比较了各种逆流、错流和模拟移动床色谱分离技术和原理,提出各种柱组合分离的方法.模拟移动床色谱能获得较高生产效率,但其设备和操作较复杂,一定程度上限制了它的应用.循环色谱是一种新颖的分离技术,这一概念最早是由Bombaugh和Biesenberger…  相似文献   

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