Comparison of chromatographic ion-exchange resins - V Strong and weak cation-exchange resins
Comparison of chromatographic ion-exchange resins - V Strong and weak cation-exchange resins
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DOI：
10.1016/j.chroma.2006.03.116
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发表时间：
2006-06-23
影响因子：
4.1
通讯作者：
Jensen, Inge Holm
中科院分区：
文献类型：
--
作者：
Staby, Arne;Jacobsen, Jan H.;Jensen, Inge Holm
比较了强阳离子交换剂和弱阳离子交换剂的一系列色谱参数，即pH依赖性、效率、结合强度、粒度分布、静态和动态容量以及扫描电子显微镜(SEM)图像。所研究的层析树脂是FractoGel EMD SO3-(M)、FractoGel EMD SE HICAP(M)、FractoGel EMD COO-(M)、MacroPrep 25S、MacroPrep High S、MacroPrep、CM、CM HyperZ和Matrex CellufineC-500。用三种蛋白进行检测：抗FVII单抗(Ig G)、抑肽酶和溶菌酶。对于等电点高于实验pH的溶菌酶和抑肽酶，pH对保留率的依赖性一般较低，但CM HyperZ的保留率随pH的升高而明显下降。对于等电点为7.5的抗FVII单抗，在pH 7.5时可与几种树脂结合。效率结果表明，随着流量的增加，板高度的依赖性增加，这是预期的趋势，而FractoGel EMD COO-的流量依赖性最高。用库尔特计数和扫描电子显微镜照片两种独立的方法测定了颗粒大小分布，结果基本一致。作为离子强度函数的阳离子交换树脂的结合强度数据取决于蛋白质，但在高盐浓度下结合和洗脱通常可以用MacroPrep树脂进行。对10%穿透时的动态容量数据和静态容量测量数据的比较表明，在层析操作过程中使用了非常不同的利用率，大约占总可用容量的25%-90%。研究了酵母发酵组分的竞争结合对抑肽酶动态结合能力的影响，发现结合能力显着下降。SepharoseM Toyota opearl 650M和陶瓷HyperD F强阳离子交换树脂和弱阳离子交换树脂被纳入本研究。具有良好纯抑肽酶活性的树脂对发酵液中的抑肽酶也有较好的吸附效果，但相对容量最高的是MacroPrep High S，其纯组分动态容量较低。本论文的结果可用于生物制药蛋白质纯化工艺开发中进一步测试的树脂的选择。(C)2006爱思唯尔B.V.保留所有权利。
Strong and weak cation-exchangers were compared for a number of chromatographic parameters, i.e. pH dependence, efficiency, binding strength, particle size distribution, static and dynamic capacity, and scanning electron microscopy (SEM) pictures. Chromatographic resins investigated were Fractogel EMD SO3- (M), Fractogel EMD SE Hicap (M), Fractogel EMD COO- (M), MacroPrep 25S, MacroPrep High S, MacroPrep, CM, CM HyperZ, and Matrex Cellufine C-500. Testing was done with three proteins: Anti-FVII Mab (IgG), aprotinin, and lysozyme. For lysozyme and aprotinin with pI above experimental pH, dependence of pH on retention was generally low, though some pronounced decrease of retention with increasing pH was observed for CM HyperZ. For Anti-FVII Mab with pI < 7.5, binding was observed on several resins at pH 7.5. Efficiency results present the expected trend of increasing dependence of plate height as a function of increasing flow rate, and the highest flow dependence was observed for Fractogel EMD COO-. Particle size distribution was determined by two independent methods, coulter counting and SEM pictures, with fair agreement. Binding strength data of cation-exchange resins as a function of ionic strength depends on the protein, but binding and elution at high salt concentration may in general be performed with MacroPrep resins. Comparison of dynamic capacity data at 10% break-through and static capacity measurements shows that a very diverse utilization of approximately 25-90% of the total available capacity is employed during chromatographic operation. The effect of competitive binding from yeast fermentation components on dynamic binding capacity of aprotinin was studied showing a significant decrease in binding capacity. Sepharose M Toyopearl 650 M, and Ceramic HyperD F strong and weak cation-exchange resins were included in this study. Resins with good pure aprotinin capacity also performed well for aprotinin in fermentation broth, but the highest relative capacity was obtained with MacroPrep High S having a fairly low pure component dynamic capacity. Results of this paper may be used in the selection of resins for further testing in biopharmaceutical protein purification process development. (c) 2006 Elsevier B.V. All rights reserved.