Linear gene fusions of antibody fragments with streptavidin can be linked to biotin labelled secondary molecules to form bispecific reagents
Linear gene fusions of antibody fragments with streptavidin can be linked to biotin labelled secondary molecules to form bispecific reagents
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抗体片段与链霉亲和素的线性基因融合可以与生物素标记的二级分子连接，形成双特异性试剂
DOI：
10.1080/15216549700203651
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发表时间：
1997
期刊：
影响因子：
4.6
通讯作者：
G. Lilley
中科院分区：
文献类型：
--
作者：
L. Pearce;G. Oddie;Gregory Coia;A. Kortt;P. Hudson;G. Lilley
单体单链抗体（scFv）片段既缺乏二价IgG或（Fab′）2片段的亲合力，又缺乏Fc结构域赋予的效应子功能。对于某些诊断或治疗应用，可能需要将这些分子与其他蛋白质、抗体、酶或肽配体连接，并且已经开发了化学或重组方法来生产许多这些交联试剂。一种方法是将抗体片段连接至链霉亲和素，其可以结合第二生物素化分子以产生更高亲和力的双功能或双特异性分子。为了证明该技术的适用性，在E.大肠杆菌中进行复性，并从6 M盐酸胍中纯化至均一。在BIAcoreTM生物传感器中的分析显示，NC10 scFv部分与固定的神经氨酸酶反应，并且核心链霉亲和素部分能够结合生物素化的抗铁蛋白Fab'，以产生结合铁蛋白的新模型双特异性试剂。从概念上讲，这种设计原理可以应用于创造有用的诊断和可能的治疗分子。
Monomeric single chain antibody (scFv) fragments lack both the avidity of the bivalent IgG, or (Fab′)2 fragment, and the effector functions conferred by the Fc domain. For certain diagnostic or therapeutic applications it may be desirable to link these molecules to other proteins, antibodies, enzymes or peptide ligands, and chemical or recombinant methods have been developed to produce many of these crosslinked reagents. One approach has been to link an antibody fragment to streptavidin which can bind a second biotinylated molecule to create a higher affinity, bifunctional or bispecific molecule. To demonstrate the applicability of this technology, an anti‐neuraminidase NC10 scFv‐streptavidin fusion was expressed in E. coli and the product was refolded and purified to homogeneity from 6 M guanidine hydrochloride. Analysis in a BIAcoreTM biosensor showed that the NC10 scFv moiety reacted with immobilised neuraminidase and that the core streptavidin moiety was able to bind biotinylated anti‐ferritin Fab' to produce a new model bispecific reagent which bound ferritin. Conceptually, this design principle can be applied to the creation of useful diagnostic and possibly therapeutic molecules.