Recombinant Escherichia coli strain produces a ZZ domain displaying biopolyester granules suitable for immunoglobulin G purification
Recombinant Escherichia coli strain produces a ZZ domain displaying biopolyester granules suitable for immunoglobulin G purification
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DOI：
10.1128/aem.01014-06
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发表时间：
2006-11-01
影响因子：
4.4
通讯作者：
Rehm, Bernd H. A.
中科院分区：
文献类型：
--
作者：
Brockelbank, Jane A.;Peters, Verena;Rehm, Bernd H. A.
将来自金黄色葡萄球菌的蛋白A的免疫球蛋白G（IgG）结合ZZ结构域融合到来自钩虫贪铜菌的聚羟基链烷酸（PHA）合酶的N末端。通过基质辅助激光解吸电离飞行时间质谱和介导的ZZ结构域展示PHA颗粒在重组大肠杆菌中的形成证实融合蛋白。使用酶联免疫吸附测定法评估分离颗粒的IgG结合能力，并且可以通过ZZ-PHA合酶的过度生产来增强。ZZ-PHA颗粒能够有效地从人血清中纯化IgG。
The immunoglobulin G (IgG) binding ZZ domain of protein A from Staphylococcus aureus was fused to the N terminus of the polyhydroxyalkanoate (PHA) synthase from Cupriavidus necator. The fusion protein was confirmed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry and mediated formation of ZZ domain-displaying PHA granules in recombinant Escherichia coli. The IgG binding capacity of isolated granules was assessed using enzyme-linked immunosorbent assay and could be enhanced by the overproduction of the ZZ-PHA synthase. ZZ-PHA granules enabled efficient purification of IgG from human serum.