摘要
Hyperimmune equine IgG is widely used as antivenom and anti-rabies agents. This article discusses a membrane based enhanced hybrid bioseparation technique for efficient and scalable purification of equine immunoglobulin G (IgG) from horse serum. This technique is an improved version of a standard hybrid bioseparation technique developed within our group earlier for fractionation of human plasma proteins (Ghosh. 2004. J Membr Sci 237: 109–117). In the presence of a high antichaotropic salt concentration, equine IgG is selectively and reversibly captured within a stirred cell membrane module from horse serum, partly due to precipitation and microfiltration, and partly due to hydrophobic interaction based membrane adsorption, while the impurities are washed out from the device. The reversibly sequestered IgG is then released by lowering the salt concentration which favor both dissolution of the precipitated IgG and desorption of the membrane bound IgG. The enhanced hybrid bioseparation technique improves the IgG recovery from the membrane module by switching from a stirring to non-stirring mode during the IgG release phase. It also reduces membrane fouling by an appropriate pH switch. The effects of operating conditions on equine IgG capture were first systematically studied. The enhanced hybrid bioseparation technique was followed by an ultrafiltration step to remove ammonium sulfate and low molecular weight impurities. The equine IgG purity obtained under optimized conditions was 88% and its recovery was over 90%, both being significantly higher than corresponding values obtained using currently used purification techniques. Biotechnol. Bioeng. 2008;99: 625–633. © 2007 Wiley Periodicals, Inc.
摘要译文
超免疫马的IgG被广泛用作抗蛇毒血清和抗狂犬病剂。本文讨论了马免疫球蛋白G（IgG抗体）从马血清高效，可扩展的净化膜基于增强型混合生物分离技术。这种技术是我们集团内制定早期人类血浆蛋白质（戈什的分级标准混合生物分离技术的改进版，2004年。ĴMembr科学237：109-117）。在高antichaotropic盐浓度的存在下，马IgG的选择性和可逆地从马血清的搅拌细胞膜模块内捕获部分原因是由于沉淀和微滤，并且部分是由于疏水基相互作用膜吸附，而杂质从设备洗出。可逆螯合的IgG然后通过降低盐的浓度有利于结合IgG膜的沉淀IgG和解吸都溶解释放。增强的混合生物分离技术改进了从膜组件IgG的回收率通过从搅拌到非搅拌模式中的IgG的释放阶段的切换。它还通过一个合适的pH值降低开关膜污染。对马IgG抗体捕获操作条件的影响，第一个系统研究。增强的混合生物分离技术，随后超滤步骤以除去硫酸铵和低分子量杂质。优化的条件下得到的抗体的马纯度为88％，其回收率可达90％以上，既比对应使用目前使用的纯化技术获得的值显著更高。生物工程。生物工程学。 2008; 99：625-633。
Lu Wang, Xinghua Sun and Raja Ghosh[*]. Purification of equine IgG using membrane based enhanced hybrid bioseparation technique: A potential method for manufacturing hyperimmune antibody[J]. Biotechnology and Bioengineering, 2008,99(3): 625-633