Hapten mediated display and pairing of recombinant antibodies accelerates assay assembly for biothreat countermeasures.
Hapten mediated display and pairing of recombinant antibodies accelerates assay assembly for biothreat countermeasures.
复制标题
半抗原介导的重组抗体的展示和配对可加速生物威胁对策的测定组装。
DOI：
10.1038/srep00807
复制
发表时间：
2012
影响因子：
4.6
通讯作者：
Hayhurst,Andrew
中科院分区：
文献类型：
--
作者：
Sherwood,LauraJ;Hayhurst,Andrew
重组抗体三明治免疫分析发展的瓶颈是配对，这需要对蛋白质进行纯化和修饰以区分捕获物和示踪物。我们开发了一个简单的配对方案，使用微升量的e。大肠渗透冲击波携带位点特异性生物素化抗体，并证明了单域抗体（sdAb）的原理证明，该抗体既是多价马尔堡病毒核蛋白的捕获者，也是示踪者。该系统还可以宿主针对7种肉毒杆菌神经毒素（BoNT）血清型的不同sdAb对，从而能够识别同源血清型。inducibesupeco -expression使sdAb群体既可以作为噬菌体繁殖，以便更多地筛选，也可以作为可溶性sdAb表达，以便在单个宿主菌株中进行筛选。当与链亲和素-g3p融合物结合时，构建了一种新型的转显示系统，以改进半合成的sdAb库，该库可用于立即免疫测定的抗埃博拉病毒sdAb，从而加快了重组抗体的发现和利用过程。
A bottle-neck in recombinant antibody sandwich immunoassay development is pairing, demanding protein purification and modification to distinguish captor from tracer. We developed a simple pairing scheme using microliter amounts ofE. coliosmotic shockates bearing site-specific biotinylated antibodies and demonstrated proof of principle with a single domain antibody (sdAb) that is both captor and tracer for polyvalentMarburgvirusnucleoprotein. The system could also host pairs of different sdAb specific for the 7 botulinum neurotoxin (BoNT) serotypes, enabling recognition of the cognate serotype. InduciblesupEco-expression enabled sdAb populations to be propagated as either phage for more panning from repertoires or expressed as soluble sdAb for screening within a single host strain. When combined with streptavidin-g3p fusions, a novel transdisplay system was formulated to retrofit a semi-synthetic sdAb library which was mined for an anti-EbolavirussdAb which was immediately immunoassay ready, thereby speeding up the recombinant antibody discovery and utilization processes.