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.

作物保护策略未来发展的一个重要目标是选择性地针对害虫物种而不是非害虫物种。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。然而，抗体相对较大的尺寸限制了组织进入和渗透到靶位点。在本提案中，我们将研究纳米抗体：抗体可变重链的单链简单多肽等价物 (1)。它们具有表位特异性，可以选择性地靶向分子并执行功能扰动。纳米抗体简化了重组表达，并提供了易于通过转基因方法引入生物系统的干扰技术。我们提出了利用秀丽隐杆线虫的实验易处理性来转化纳米抗体技术的实验。1。秀丽隐杆线虫的优势。线虫是食细菌动物，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，都优先采用基于喂养的方法 (2,3)。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因素，其破坏在定量行为测定中进行评分。这些表型反映了已建立的害虫化学缓解方法中所发生的破坏。此外，线虫允许系统的遗传操作来表达带有人工标签的候选目标分子。针对此类标签的商业纳米抗体可以探究新靶标或途径的潜力。2．支持纳米抗体研究的化学生物学。完全体外方法可以识别候选纳米抗体和编码 cDNA。工程靶点中人工标签的商业纳米抗体将用于启动工作。控制功能或蛋白质-蛋白质相互作用的已知拓扑结构和关键域将优化功能干扰纳米抗体候选表位的筛选 (5).3。研究和调节纳米抗体的分布。一个重要的目标是验证纳米抗体作为可用于作物保护的生物制剂。蠕虫提供了一个机会来探索在复杂生物体的背景下如何接近目标。我们将测试设计用于表达表位识别的纳米抗体，无论是否带有细胞和组织穿透标签 (6)。通过喂食天然或修饰实体，这些短段的工程序列已发现纳米抗体或修饰纳米抗体对生物体的渗透比较 (7)。秀丽隐杆线虫模型的开发将突出用于在更易遗传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穿过细胞边界方面具有卓越的价值。