二聚体双特异性微型抗体以亲和力结合两种特异性。

A dimeric bispecific miniantibody combines two specificities with avidity.

作者信息

Müller K M, Arndt K M, Plückthun A

机构信息

Biochemisches Institut, Universität Zürich, Switzerland.

出版信息

FEBS Lett. 1998 Jul 31;432(1-2):45-9. doi: 10.1016/s0014-5793(98)00829-1.

DOI:10.1016/s0014-5793(98)00829-1

PMID:9710248

Abstract

Bispecific antibodies extend the capabilities of nature and might be applied in immunotherapy and biotechnology. By fusing the gene of a single-chain Fv (scFv) fragment to a helical dimerization domain, followed by a second scFv fragment of different specificity, we were able to express a functional protein in E. coli, which is bispecific and has two valencies for each specificity. The dimeric bispecific (DiBi) miniantibody preserves the natural avidity of antibodies in a very small-sized molecule of only 120 kDa. The generality of the principle was shown with a scFv fragment binding the EGF-receptor (named scFv 425) in three combinations with scFv fragments either directed against CD2 (ACID2.M1), phosphorylcholine (McPC603) or fluorescein (FITC-E2). Binding was analyzed by sandwich surface plasmon resonance biosensor (BIAcore) measurements.

摘要

双特异性抗体拓展了自然的能力，可应用于免疫治疗和生物技术领域。通过将单链Fv（scFv）片段的基因与一个螺旋二聚化结构域融合，随后再接上一个具有不同特异性的第二个scFv片段，我们得以在大肠杆菌中表达一种功能性蛋白，该蛋白具有双特异性，且每种特异性都有两个结合价。二聚体双特异性（DiBi）微型抗体在仅120 kDa的非常小的分子中保留了抗体的天然亲和力。用与表皮生长因子受体结合的scFv片段（命名为scFv 425）与分别针对CD2（ACID2.M1）、磷酸胆碱（McPC603）或荧光素（FITC-E2）的scFv片段的三种组合展示了该原理的通用性。通过夹心表面等离子体共振生物传感器（BIAcore）测量分析结合情况。