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Fermentation Microbiology and Biotechnology E.M.T. El-Mansi and C.F.A. Bryce (Eds.) 发酵微生物学和生物技术E.M.T. El-Mansi和C.F.A. Bryce(编)
Bioseparation
Pub Date : 2000-07-01 DOI: 10.1023/A:1008192103524
J. Kennedy, N. Turan
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引用次数: 1
Superporous agarose monoliths as mini-reactors in flow injection systems 超多孔琼脂糖单体在流动注射系统中的微型反应器
Bioseparation
Pub Date : 2000-07-01 DOI: 10.1023/A:1008117827057
Madayi P. Nandakumar, E. Pålsson, P. Gustavsson, P. Larsson, B. Mattiasson
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引用次数: 20
Biological Process Engineering A. T. Johnson 生物过程工程
Bioseparation
Pub Date : 2000-07-01 DOI: 10.1023/A:1008105524174
J. Kennedy, M. Thorley
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引用次数: 5
Chromatography and Capillary Electrophoresis in Food Analysis H. Sorensen, S. Sorensen, C.Bjergegaard and S. Michaelsen 色谱和毛细管电泳在食品分析中的应用
Bioseparation
Pub Date : 2000-07-01 DOI: 10.1023/A:1008182232750
J. Kennedy, Lorraine A. Quinton
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引用次数: 0
Freeze-Drying/ Lyophilization of Pharmaceutical and Biological Products, Louis Rey and Joan C. May 制药和生物制品的冷冻干燥/冻干，Louis Rey和Joan C. May
Bioseparation
Pub Date : 2000-03-01 DOI: 10.1023/A:1008125212856
J. Kennedy, N. Turan
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引用次数: 2
Immobilized Biomolecules in Analysis: A Practical Approach, Tony Cass & Frances S. Ligler (Eds.) 固定化生物分子分析:一种实用的方法，托尼卡斯和弗朗西斯S.利格勒(编)
Bioseparation
Pub Date : 2000-03-01 DOI: 10.1023/A:1008192009124
J. Kennedy, N. Turan
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引用次数: 4
HPLC of Macromolecules, 2 nd Edition, R.W.A. Oliver (Editor) 大分子高效液相色谱，第2版，R.W.A. Oliver(编辑)
Bioseparation
Pub Date : 2000-03-01 DOI: 10.1023/A:1008126231841
J. Kennedy, Lorraine A. Quinton
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引用次数: 0
Two-dimensional polyacrylamide gel electrophoresis of the protease SP220K, a renal cell carcinoma marker. 肾细胞癌标志物蛋白酶SP220K的双向聚丙烯酰胺凝胶电泳。
Bioseparation
Pub Date : 2000-01-01 DOI: 10.1023/a:1008198521231
M Starita-Geribaldi, S Thaon-Scarzello, M Le Blanc, E Van Obberghen, C Poustis-Delpont

The present study analyses, by two-dimensional polyacrylamide gel electrophoresis, the protease SP220K isolated from renal cell carcinoma. The pure molecule is separated using either immobilized pH gradient or isoelectric focusing in conventional carrier ampholyte in the first dimension. Some interactions with the acrylamide matrix in isoelectric focusing are discussed. The results demonstrate that two-dimensional gel electrophoresis performed with enriched media such as basolateral membranes, allows the detection of the protease. In addition, the non detection of the molecule up to now by this methodology can be explained by the high tendency of oligomerization of SP220K. Effectively the high molecular weight form of the molecule of 220 kDa is favoured in two-dimensional gel electrophoresis over monomeric forms which are better detected in SDS PAGE. This was confirmed by immunostaining performed with an antiserum to SP220K produced by nitrocellulose-bound antigen.

本研究采用双向聚丙烯酰胺凝胶电泳法对肾细胞癌中分离的蛋白酶SP220K进行了分析。采用固定化pH梯度或等电聚焦方法在常规载体两性电解质中一维分离纯分子。讨论了等电聚焦过程中与丙烯酰胺基体的相互作用。结果表明，二维凝胶电泳进行富集介质，如基底外侧膜，允许检测蛋白酶。此外，该方法至今未检测到该分子，可以用SP220K的高寡聚倾向来解释。在二维凝胶电泳中，220 kDa分子的高分子量形式比在SDS PAGE中更好地检测到的单体形式更受青睐。用硝基纤维素结合抗原产生的SP220K抗血清进行免疫染色证实了这一点。
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引用次数: 3
Peptide fusion tags with tryptophan and charged residues for control of protein partitioning in PEG-potassium phosphate aqueous two-phase systems. 与色氨酸和带电荷残基的肽融合标签在peg -磷酸钾两水相体系中用于控制蛋白质分配。
Bioseparation
Pub Date : 2000-01-01 DOI: 10.1023/a:1008182711385
K Berggren, F Tjerneld, A Veide

A partition study with peptides and recombinant proteins in poly(ethylene glycol)4000-potassium phosphate aqueous two-phase systems has been performed. The aim was to study to what extent the insertion of charged residues could affect protein partition in addition to the already observed effects of tryptophan residues. The model proteins used are based on a staphylococcal protein A derivative, Z, and modified by the insertion of peptide tags close to the C-terminus. The tags differed with respect to their content of both Trp, negatively (Asp) and positively charged (Lys) amino acid residues. The same partitioning trends were observed for the peptides and fusion proteins. The effect of Trp residues was to direct the partitioning towards the PEG phase. The insertion of two negatively charged (Asp) residues into a Trp4-tag enhanced the partition towards the PEG phase even more. The introduction of positively charged (Lys) residues in addition to Trp residues, on the other hand, pulled the peptide or protein towards the potassium phosphate phase. The partitioning of peptides gave a good qualitative picture of the effect of the peptide on partitioning when fused to the protein. The efficiencies of the tags were calculated based on partitioning of tags and fusion proteins, and tag efficiencies generally varied between 60 and 85%.

多肽和重组蛋白在聚乙二醇4000-磷酸钾双水相体系中的分配研究已经完成。目的是研究除了色氨酸残基已经观察到的影响外，带电残基的插入在多大程度上可以影响蛋白质的分配。所使用的模型蛋白基于葡萄球菌蛋白a衍生物Z，并通过插入靠近c端的肽标签进行修饰。这些标签在色氨酸、带负电的(Asp)和带正电的(Lys)氨基酸残基的含量方面存在差异。在多肽和融合蛋白中观察到相同的分裂趋势。色氨酸残基的作用是直接向PEG相分配。在trp4标签中插入两个带负电荷的(Asp)残基进一步增强了向PEG相的分裂。另一方面，除了色氨酸残基外，引入带正电荷(Lys)残基将肽或蛋白质拉向磷酸钾相。多肽的分配给了一个很好的定性的图片肽对分配的影响，当融合到蛋白质。标签的效率是根据标签和融合蛋白的分配来计算的，标签效率一般在60%到85%之间变化。
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引用次数: 17
Bovine serum albumin-hemoglobin fractionation: significance of ultrafiltration system and feed solution characteristics. 牛血清白蛋白-血红蛋白分离:超滤系统和饲料溶液特性的意义。
Bioseparation
Pub Date : 2000-01-01 DOI: 10.1023/a:1008194300403
R Shukla, M Balakrishnan, G P Agarwal

This work investigates the fractionation of similar molecular weight proteins bovine serum albumin (69 kD) and bovine hemoglobin (67 kD) by ultrafiltration. Three different membranes, viz. regenerated cellulose, poly(sulfone) and surface modified poly(acrylonitrile), each with a nominal molecular cutoff rating of 100 kD, were examined. The experiments were conducted in dead end, crossflow and vortex flow filtration modes and the separation was studied as a function of feed pH and ionic strength. Under similar system hydrodynamics, the surface modified poly(acrylonitrile) membrane displayed the highest resolution with minimum membrane fouling. The separation could be improved further by operating at low applied pressure (40 kPa) and high mass transfer (> 20 x 10(-6) m/s) in a vortex flow module. Under these conditions, the highest separation factor of 40 was obtained at the pI of hemoglobin.

本文研究了用超滤法分离分子量相近的牛血清白蛋白(69 kD)和牛血红蛋白(67 kD)。研究了三种不同的膜，即再生纤维素、聚砜和表面改性聚丙烯腈，每种膜的标称分子截止额定值为100 kD。实验分别在死角、横流和涡旋三种过滤模式下进行，并研究了进料pH和离子强度对分离效果的影响。在相似的系统流体力学条件下，表面改性聚丙烯腈膜具有最高的分辨率和最小的膜污染。在低施加压力(40 kPa)和高传质(> 20 x 10(-6) m/s)的涡流模块中，分离可以进一步改善。在此条件下，在血红蛋白的pI处获得了最高的分离因子40。
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引用次数: 34
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