Grant S D, Cupit P M, Learmonth D, Byrne F R, Graham B M, Porter A J, Harris W J
Department of Molecular and Cell Biology, University of Aberdeen, Marischal College, UK.
J Hematother. 1995 Oct;4(5):383-8. doi: 10.1089/scd.1.1995.4.383.
The technology of humanization of rodent antibodies has opened the way for a broad range of therapeutic antibodies with very low immunogenicity, which are, therefore, suitable for repeated dosing. Such intact antibodies have extended serum half-lives and biodistribution profiles very similar to human antibodies. For some applications, however, the ideal therapeutic should have reduced serum half-life and altered biodistribution patterns typical of antibody fragments, such as Fab or single chain Fv. Bispecific antibody fragments offer exciting additional therapeutic possibilities, but their successful manufacture and purification on a large scale require the development of new methods. Antibody fragments often assemble in Escherichia coli as monovalent fragments with reduced affinities. We describe the spontaneous assembly of bivalent antibody fragments in E. coli and methods of purification that yield either bivalent or monovalent molecules as required.
啮齿动物抗体人源化技术为一系列免疫原性极低的治疗性抗体开辟了道路，因此适用于重复给药。这类完整抗体具有延长的血清半衰期，其生物分布特征与人类抗体非常相似。然而，对于某些应用而言，理想的治疗药物应具有缩短的血清半衰期以及抗体片段（如Fab或单链Fv）典型的改变的生物分布模式。双特异性抗体片段提供了令人兴奋的额外治疗可能性，但它们的大规模成功生产和纯化需要开发新方法。抗体片段在大肠杆菌中常组装成亲和力降低的单价片段。我们描述了二价抗体片段在大肠杆菌中的自发组装以及根据需要产生二价或单价分子的纯化方法。
