Peptide-based tool for the rapid isolation of quiescent monocytes from peripheral blood

用于从外周血中快速分离静态单核细胞的基于肽的工具

• 批准号: 9340352
• 负责人: Martyn Darby
• 金额: $ 30.88万
• 依托单位: AFFINERGY, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-09 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9340352
• 关键词: Address; Adherence; Amination; Amines; Antibodies; Bacteriophages; Binding; Blood; Blood Cells; Blood Platelets; C-terminal; CD14 gene; CD19 gene; Cancer Vaccines; Cell Differentiation process; Cell surface; Cells; Chemistry; Clinic; Clinical; Dendritic Cell Therapy; Dendritic Cell Vaccine; Dendritic Cells; Development; Erythrocytes; Family; Generations; Goals; Hematopoietic stem cells; Hour; Immunotherapeutic agent; In Vitro; Interleukin-2; Life; Lymphocyte; Lymphocyte Activation; Magnetic Bead Technology; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Methods; Mononuclear; Peptides; Peripheral; Peripheral Blood Mononuclear Cell; Phage Display; Phase; Phenotype; Plasma; Plasticizers; Population; Protocols documentation; Recovery; Series; Source; Sulfhydryl Compounds; T-Lymphocyte; Technical Expertise; Techniques; Technology; Testing; Time; Vaccination; Vaccines; Whole Blood; base; cadmium ion; cancer cell; cancer immunotherapy; cancer therapy; cell type; clinical development; cost; cost efficient; cross reactivity; cytokine; density; granulocyte; large scale production; magnetic beads; manufacturing scale-up; monocyte; peripheral blood; precursor cell; prototype; rapid technique; tool; tumor

SUMMARY/ABSTRACT Dendritic cell (DC) therapy represents a new and promising immunotherapeutic approach for treatment of advanced stage cancers, with DC vaccines already approved for use in advanced prostate cancer, and more than 30 DC-based vaccines in the clinical development pipeline for other cancers. In particular, the US market for cancer vaccines (including DC-based vaccines) is set to expand tremendously at a CAGR of 78.1% from $20 million in 2010 to $2 billion by 2018. This highlights the need to develop clinical grade technologies for collecting DC precursor cells and generating DCs for cancer vaccination. Typically, DCs are differentiated from monocytes, which are isolated from peripheral blood mononuclear cells using different methods including plastic adherence, counter-flow elutriation, and immunomagnetic selection or depletion of cells. Of these techniques, immunomagnetic selection of CD14+ monocytes is the most commonly employed technique in the clinic. While this technique results in a relatively pure population of monocytes as compared to the other techniques, the recovery/yield of monocytes is low. Also, the use of antibodies to purify monocytes makes it cost-prohibitive for large-scale production of monocytes. In addition, all the above mentioned techniques are unable to isolate monocytes from whole blood, and require a density-gradient step prior to isolation of monocytes, thus increasing costs, time and technical skill involved. Thus, there is an unmet need to develop a simple, rapid and efficient method capable of isolating a highly pure and quiescent population of monocytes directly from whole blood. Affinergy has identified a family of peptides that bind specifically to monocytes, while not binding to the other blood cells. When conjugated to magnetic beads, these peptides can directly isolate monocytes from whole blood, and result in high recovery and purity, without any additional processing steps. At the conclusion of Phase I, we will have established a rapid and efficient method for purifying monocytes from whole blood and demonstrated that the enriched monocytes are capable of differentiating into DCs. In Phase II, we will optimize large-scale production of peptide-conjugated beads, establish shelf-life and storage conditions and demonstrate the feasibility of our technique to purify monocytes at a larger scale.

摘要/摘要 树突状细胞（DC）疗法代表了一种新的、有前途的免疫治疗方法，用于治疗 晚期癌症，DC 疫苗已被批准用于晚期前列腺癌等 超过 30 种基于 DC 的疫苗正在针对其他癌症进行临床开发。尤其是美国市场 癌症疫苗（包括基于 DC 的疫苗）将以 78.1% 的复合年增长率大幅增长 2010 年的 2000 万美元到 2018 年将达到 20 亿美元。这凸显了开发临床级技术的必要性 收集 DC 前体细胞并生成用于癌症疫苗接种的 DC。通常，DC 与 单核细胞，使用不同的方法从外周血单核细胞中分离出来，包括 塑料粘附、逆流淘析以及免疫磁性选择或细胞耗竭。其中 技术中，CD14+单核细胞的免疫磁选是最常用的技术 诊所。虽然与其他技术相比，该技术产生了相对纯净的单核细胞群 技术，单核细胞的回收率/产量较低。此外，使用抗体纯化单核细胞使其 大规模生产单核细胞的成本过高。此外，上述所有技术都是 无法从全血中分离单核细胞，并且在分离之前需要密度梯度步骤 单核细胞，从而增加了成本、时间和所涉及的技术技能。因此，存在未满足的开发需求 一种简单、快速和有效的方法，能够分离高纯度和静止的群体 直接来自全血的单核细胞。 Affinergy 已鉴定出一系列能够特异性结合的肽 单核细胞，而不与其他血细胞结合。当与磁珠结合时，这些肽可以 直接从全血中分离单核细胞，回收率和纯度高，无需任何额外 处理步骤。在第一阶段结束时，我们将建立一种快速有效的方法 从全血中纯化单核细胞，并证明富集的单核细胞能够 分化为DC。在第二阶段，我们将优化肽缀合珠的大规模生产， 建立保质期和储存条件并证明我们纯化单核细胞技术的可行性 在更大的范围内。