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首页> 外文期刊>Biotechnology Progress >A novel magnetic affinity support for protein adsorption and purification
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A novel magnetic affinity support for protein adsorption and purification

机译：一种新型的磁性亲和支持物，用于蛋白质吸附和纯化

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A novel magnetic support was prepared by an oxidization-precipitation method with poly(vinyl alcohol) (PVA) as the entrapment material. Transmission electron microscopy indicated that the magnetic particles had a core-shell structure, containing many nanometer-sized magnetic cores stabilized by the cross-linked PVA. The particles showed a high magnetic responsiveness in magnetic field, and no aggregation of the particles was observed after the particles had been treated in the magnetic field. These facts indicated that the particles were superparamagnetic. Cibacron blue 3GA (CB) was coupled to the particles to prepare a magnetic affinity support (MAS) for protein adsorption. Lysozyme was used as a model protein to test the adsorption properties of the MAS. The adsorption equilibrium of lysozyme to the MAS was described by the Langmuir-type isotherm. The capacity for lysozyme adsorption was more than 70 mg/g MAS (wet weight) at a relatively low CB coupling density (3-5 mu mol/g). In addition, 1.0 M NaCl solution could be used to dissociate the adsorbed lysozyme. Finally, the MAS was recycled for the purification of alcohol dehydrogenase (ADH) from clarified yeast homogenates. Under proper conditions, the magnetic separation yielded over 5-fold purification of the enzyme with 60% recovery of the enzyme activity.
机译：以聚乙烯醇（PVA）为包埋材料，通过氧化沉淀法制备了新型磁性载体。透射电子显微镜表明，磁性颗粒具有核-壳结构，包含许多通过交联的PVA稳定的纳米级磁性核。颗粒在磁场中显示出高的磁响应性，并且在将颗粒在磁场中处理之后未观察到颗粒的聚集。这些事实表明颗粒是超顺磁性的。将烟蓝3GA（CB）与颗粒偶联，以制备用于蛋白质吸附的磁性亲和载体（MAS）。溶菌酶被用作模型蛋白以测试MAS的吸附特性。兰格缪尔型等温线描述了溶菌酶对MAS的吸附平衡。在相对较低的CB耦合密度（3-5μmol / g）下，溶菌酶的吸附能力大于70 mg / g MAS（湿重）。另外，可以使用1.0 M NaCl溶液解离吸附的溶菌酶。最后，将MAS再循环以从澄清的酵母匀浆中纯化乙醇脱氢酶（ADH）。在适当的条件下，磁分离产生的酶纯度提高了5倍以上，酶活性恢复了60％。

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