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 基因疗法和噬菌体疗法的领域正在迅速扩展，并显示出令人鼓舞的 分别治疗遗传疾病和抗生素耐药细菌感染的潜力。 为了满足这些需求，改善高吞吐量和高亲和力病毒纯化方法将是 必需的。当前的高通量病毒纯化方法主要由基于树脂的基于树脂组成 色谱与多个过滤步骤结合。基于色谱的方法通常缺乏 特定病毒靶标的特异性。例如，常用的离子交换树脂 无法将共享与病毒颗粒的表面净电荷相似的污染物分离。 选择混合病毒品种，例如不同血清型的病毒 色谱法。基于亲和力的树脂色谱产品靶向病毒载体 存在但昂贵或不适合大规模纯化。此外，它们涉及 将约束力实体耦合到树脂，这表现出几个缺点，包括浸出 来自色谱支撑或配体与病毒颗粒共洗脱的亲和力配体。 因此，需要改进的病毒载体的简单，快速和可扩展的纯化的方法 服务小规模的研究社区和大规模的工业生产。邦德威尔 Technologies提议开发一种新型的纯化系统，以用于大分子，例如 病毒载体。我们功能化的生物材料的独特品质提供了巨大的多功能性 并解决了小型和大规​​模病毒纯化的当前局限性。在此期间 提议的第一阶段努力，我们将首先通过能够的实体功能化生物材料 专门识别病毒靶标。然后，我们将测试生物材料的结合能力 病毒靶标。最后，我们将准备生物材料用作膜并测试其能力 结合具有不同复杂性起始材料的病毒靶标（例如，用细胞碎片 展示）。