Extraction of monoclonal antibodies (IgG1) using anionic and anionic/nonionic reverse micelles.
Extraction of monoclonal antibodies (IgG1) using anionic and anionic/nonionic reverse micelles.
复制标题
使用阴离子和阴离子/非离子反胶束提取单克隆抗体 (IgG1)。
DOI：
10.1002/btpr.453
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发表时间：
2010
影响因子：
2.9
通讯作者：
George DA
中科院分区：
文献类型：
--
作者：
George DA
基于亲和层析的抗体生产纯化方案正试图跟上细胞培养表达水平的增长，许多当前的研究计划都集中在寻找用于纯化单克隆抗体（mab）的层析的替代品。在本文中，我们研究了一种基于液-液萃取的替代分离技术——反胶束萃取。我们使用阴离子表面活性剂（AOT）和阴离子（AOT）和非离子表面活性剂（Brij‐30,Tween‐85,Span‐85）的组合（以异辛烷为溶剂体系）的反胶束提取单抗（IgG1）。研究了水相pH、阳离子浓度、表面活性剂种类和浓度等参数对IgG1提取效率的影响。使用AOT/异辛烷反胶束体系，我们可以获得良好的IgG1整体提取（在80 - 90%之间），但在提取结束时，通过使用其与亲和色谱柱的结合作为活性的替代测量，IgG1的生物活性只能恢复30%。由于阴离子表面活性剂被怀疑是导致活性降低的原因之一，我们决定将非离子表面活性剂与阴离子表面活性剂联合使用，研究其对提取效率和生物活性的影响。AOT/Brij‐30/异辛烷反胶束体系的提取效果最好，总提取率达到90%以上，总活性回收率达到59%。AOT/吐温- 85/异辛烷反胶束体系的总提取率在75 - 80%之间，总活性回收率约为40-45%。结果表明，不同的表面活性剂组合体系可以显著提高IgG1的活性回收率，如果能进一步提高IgG1的回收率，该技术在单克隆抗体的下游纯化中具有相当大的前景。©2010美国化学工程师学会生物技术。掠夺。, 2010年
Purification schemes for antibody production based on affinity chromatography are trying to keep pace with increases in cell culture expression levels and many current research initiatives are focused on finding alternatives to chromatography for the purification of Monoclonal antibodies (MAbs). In this article, we have investigated an alternative separation technique based on liquid–liquid extraction called the reverse micellar extraction. We extracted MAb (IgG1) using reverse micelles of an anionic surfactant, sodium bis 2‐ethyl‐hexyl sulfosuccinate (AOT) and a combination of anionic (AOT) and nonionic surfactants (Brij‐30, Tween‐85, Span‐85) using isooctane as the solvent system. The extraction efficiency of IgG1 was studied by varying parameters, such as pH of the aqueous phase, cation concentration, and type and surfactant concentration. Using the AOT/Isooctane reverse micellar system, we could achieve good overall extraction of IgG1 (between 80 and 90%), but only 30% of the bioactivity of IgG1 could be recovered at the end of the extraction by using its binding to affinity chromatography columns as a surrogate measure of activity. As anionic surfactants were suspected as being one of the reasons for the reduced activity, we decided to combine a nonionic surfactant with an anionic surfactant and then study its effect on the extraction efficiency and bioactivity. The best results were obtained using an AOT/Brij‐30/Isooctane reverse micellar system, which gave an overall extraction above 90 and 59% overall activity recovery. An AOT/Tween‐85/Isooctane reverse micellar system gave an overall extraction of between 75 and 80% and overall activity recovery of around 40–45%. The results showed that the activity recovery of IgG1 can be significantly enhanced using different surfactant combination systems, and if the recovery of IgG1 can be further enhanced, the technique shows considerable promise for the downstream purification of MAbs. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010