Nanobody technology: feeding target authentication and mitigation strategies in crop protection.

纳米抗体技术：作物保护中的饲喂目标验证和缓解策略。

• 批准号: 2881464
• 金额: --
• 依托单位: University of Southampton
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2881464
• 关键词: Nanobody; technology; feeding; target; authentication

An important goal for the future development of crop protection strategies is selective targeting of pest species over non-pest species. Biologics based on protein architectures offer a potential route. The development of antibody technologies that disrupt molecules is an attractive approach. Conventional antibodies are oligomeric proteins with complex biogenesis. However, the relatively large size of antibodies restricts tissue access and penetration to target sites. In this proposal, we will investigate nanobodies: single chain simple polypeptide equivalents of the antibody variable heavy chain (1). These have epitope specificity to selectively target molecules and execute functional perturbation. The nanobodies have simplified recombinant expression and provide interfering technologies readily introduced into biological systems by transgenic approaches. We propose experiments designed to translate nanobody technology by taking advantage of the experimental tractability of C. elegans.1. Advantaged by C.elegans. C. elegans are bacterivores advantaging feeding based approaches whether free recombinant nanobodies or bacteria expressing nanobodies initiate reagent delivery (2,3). We will use established phenotypes that have well known molecular determinants whose disruption is scored in quantitative behavioural assays. These phenotypes mirror disruptions played out in established chemical mitigation of pests.Furthermore, C. elegans allows the genetic manipulation of the system to express candidate target molecules with artificial tags. Commercial nanobodies against such tags can interrogate the potential of novel targets or pathways.2. Chemical Biology to support nanobody investigation. Fully in-vitro approaches can identify candidate nanobodies and encoding cDNAs. Commercial nanobodies to artificial tags in engineered targets will be used to initiate work. The known topology and critical domains that control function or protein-protein interaction will optimize the screening of candidate epitopes for function perturbing nanobodies (5).3. Investigating and modulating nanobody distribution. An important goal is validation of the nanobodies as biologics that can be used in crop protection.The worm provides an opportunity to probe how accessible targets are in context of a complex organism. We will test nanobodies engineered to express epitope recognition with and without a cell and tissue penetrating tags (6). These short stretches of engineered sequence have found compare penetration of nanobodies or modified nanobodies into the organism by feeding with native or modified entity (7). Development in the model C. elegans will highlight reagents and protocols for further testing in more genetically tractable pest species. excellent value in allowing cell impermeable entities to gain passage across cellular boundaries.

未来制定作物保护战略的一个重要目标是选择性地针对有害物种而不是非有害物种。基于蛋白质结构的生物制剂提供了一条潜在的途径。开发破坏分子的抗体技术是一种有吸引力的方法。常规抗体是具有复杂生物发生的寡聚蛋白质。然而，相对大尺寸的抗体限制了组织进入和穿透靶位点。在该提案中，我们将研究纳米抗体：抗体可变重链的单链简单多肽等同物（1）。这些具有选择性靶向分子和执行功能扰动的表位特异性。所述纳米抗体具有简化的重组表达，并提供通过转基因方法容易引入生物系统的干扰技术。我们提出的实验设计，以翻译纳米抗体技术，利用实验的易处理性的C。优雅。由C. elegans。C.无论是游离重组纳米抗体还是表达纳米抗体的细菌启动试剂递送（2，3），秀丽线虫都是食细菌动物，其基于摄食的方法。我们将使用已建立的表型，这些表型具有众所周知的分子决定簇，其破坏在定量行为测定中进行评分。这些表型反映了在既定的化学减轻害虫中发生的破坏。elegans允许对系统进行遗传操作以表达具有人工标签的候选靶分子。针对这些标签的商业纳米抗体可以询问新靶点或途径的潜力。化学生物学支持纳米抗体研究。完全体外方法可以鉴定候选纳米抗体和编码cDNA。商业纳米抗体到工程目标中的人工标签将用于启动工作。已知的拓扑结构和控制功能或蛋白质-蛋白质相互作用的关键结构域将优化功能干扰纳米抗体的候选表位的筛选（5）。研究和调节纳米抗体分布。一个重要的目标是验证纳米抗体作为生物制剂可用于作物保护。蠕虫提供了一个机会，以探索如何在复杂的有机体的背景下访问的目标。我们将测试纳米抗体工程表达表位识别与和没有细胞和组织穿透标签（6）。已经发现这些短的工程化序列的延伸通过用天然或修饰的实体喂养来比较纳米抗体或修饰的纳米抗体对生物体的渗透（7）。模型C的发展。elegans将重点介绍在遗传上更易处理的害虫物种中进行进一步测试的试剂和方案。在允许细胞不可渗透的实体获得穿过细胞边界的通道方面具有优异的价值。