Novel, Targeted Method for Bacteriophage Purification

噬菌体纯化的新型靶向方法

• 批准号: 10698983
• 负责人: Christi Parham
• 金额: $ 27.54万
• 依托单位: BONDWELL TECHNOLOGIES LP
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-10 至 2025-04-09
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698983
• 关键词: Address; Adenoviruses; Affinity; Air; Antibodies; Bacterial Antibiotic Resistance; Bacterial Infections; Bacteriophages; Binding; Biocompatible Materials; Buffers; Characteristics; Charge; Chimeric Proteins; Chromatography; Communities; Consumption; Coupled; Coupling; Disadvantaged; Drosophila Proteins; Electrospinning; Ensure; Feasibility Studies; Fiber; Film; Filtration; Gene Transduction Agent; Genetic Diseases; Housing; Ion Exchange Resins; Ligands; Mechanics; Membrane; Methods; Molecular Biology Techniques; Phase; Plant Resins; Process; Property; Proteins; Proxy; Recipe; Research; Serotyping; Specificity; Surface; Syringes; System; Tandem Repeat Sequences; Technology; Testing; Therapeutic; Time; Viral; Viral Vector; Virion; Virus; Viscosity; Water; Work; cost; covalent bond; crosslink; dityrosine; gene therapy; improved; industrial production; innovation; monomer; novel; protein purification; success

PROJECT SUMMARY The fields of gene therapy and bacteriophage therapy are rapidly expanding and show promising potential for treating genetic disorders and antibiotic resistant bacterial infections, respectively. To meet these needs, improved high throughput and high affinity viral purification methods will be required. Current high throughput viral purification methods mainly consist of resin-based chromatography coupled with multiple filtration steps. Chromatography-based methods often lack specificity for the particular virus target. For example, the commonly used ion exchange resins are not able to separate contaminants sharing similar surface net charges to the virus particles. Selection of mixed virus varieties such as viruses of different serotypes cannot be achieved with chromatography methods. Affinity-based resin chromatography products targeting viral vectors exist but are expensive or not suitable for large-scale purification. Additionally, they involve coupling a binding entity to resin, which presents several disadvantages, including leaching of the affinity ligand from the chromatography support or the ligand co-eluting with the virus particles. Thus, improved methods for simple, rapid, and scalable purification of viral vectors are needed to serve both the small-scale research community and large-scale industrial production. Bondwell Technologies proposes to develop a novel purification system for large biomolecules, such as viral vectors. The unique qualities of our functionalized biomaterial provide tremendous versatility and addresses the current limitations of small- and large-scale viral purification. During this proposed Phase I effort, we will first functionalize our biomaterials with an entity capable of specifically recognizing a viral target. We will then test the biomaterial for their ability to bind the viral target. Finally, we will prepare our biomaterial to be used as a membrane and test its ability to bind a viral target with different complexities of starting material (e.g., with cellular debris present).

项目摘要 基因治疗和噬菌体治疗的领域正在迅速扩大，并显示出良好的前景 分别用于治疗遗传性疾病和抗生素耐药性细菌感染的潜力。 为了满足这些需求，将开发改进的高通量和高亲和力病毒纯化方法。 必需的.目前的高通量病毒纯化方法主要由基于树脂的纯化方法组成。 通过色谱法与多个过滤步骤偶联。基于色谱的方法往往缺乏 对特定病毒目标的特异性。例如，常用的离子交换树脂 不能分离与病毒颗粒共享类似表面净电荷的污染物。 混合病毒品种如不同血清型的病毒的选择不能通过以下方法实现： 色谱法靶向病毒载体的亲和树脂层析产品 存在但昂贵或不适合大规模纯化。此外，它们涉及 将结合实体偶联到树脂，这存在几个缺点，包括 来自层析支持物的亲和配体或与病毒颗粒共洗脱的配体。 因此，需要用于简单、快速和可规模化纯化病毒载体的改进方法， 服务于小规模研究社区和大规模工业生产。邦威 技术提出开发一种用于大生物分子的新型纯化系统， 病毒载体我们的功能化生物材料的独特品质提供了巨大的多功能性 并解决了目前小规模和大规模病毒纯化的局限性。在此 在第一阶段的工作中，我们将首先用一种能够 特异性识别病毒靶点。然后，我们将测试生物材料的结合能力， 病毒目标最后，我们将准备我们的生物材料作为膜使用，并测试其能力 为了结合具有不同复杂性的起始材料的病毒靶（例如，细胞碎片 present）。