Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528
Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528
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DOI：
10.1093/protein/gzn037
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发表时间：
2008-10-01
影响因子：
2.4
通讯作者：
Kumagai, Izumi
中科院分区：
文献类型：
--
作者：
Asano, Ryutaro;Sone, Yukiko;Kumagai, Izumi
我们报告了体外重折叠在制备具有表皮生长因子受体的单态hEx 3双特异性双抗体中的实用性，并且从大肠杆菌中的不溶性聚集体重定向CD 3。同源可变重链和轻链之间的适当相互作用导致形成双抗体形式的功能性hEx 3异二聚体，而不是无活性的同源二聚体。发现重折叠的hEx 3表现出与在哺乳动物分泌系统中制备的hEx 3和单链hEx 3（hEx 3-scDb）几乎相当的活性。我们建议，从细菌不溶性材料的hEx 3的制备通过体外复性将是有用的工业规模生产的双抗体，其潜在的临床应用研究。
We report the utility of in vitro refolding in the preparation of monomorphous hEx3 bispecific diabodies with epidermal growth factor receptor and CD3 retargeting from insoluble aggregates in Escherichia coli. Appropriate interaction between cognate variable heavy and light chains led to the formation of functional hEx3 heterodimers in a diabody format rather than inactive homodimers. The refolded hEx3 was found to exhibit almost the equivalent activity to the hEx3 and single-chain hEx3 ( hEx3-scDb) prepared in a mammalian secretion system. We suggest that the preparation of hEx3 from bacterial insoluble material by means of in vitro refolding would be useful for industrial-scale production of the diabody for its potential use in clinical studies.