Secretomes captured in vivo via ultrafiltration probes

通过超滤探针在体内捕获分泌物

• 批准号: 7283959
• 负责人: CHUN-MING HUANG
• 金额: $ 16.82万
• 依托单位: VETERANS MEDICAL RESEARCH FDN/SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2009-08-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7283959
• 关键词: Address; Animal Model; Animals; Antibodies; Biological; Biological Models; Blood capillaries; Bypass; Cells; Coupling; Cultured Tumor Cells; Data; Doctor of Philosophy; Enzyme-Linked Immunosorbent Assay; Fiber; Figs - dietary; Glucose; Goals; Immunosuppression; Implant; In Vitro; Ions; Laboratories; Life; Liquid substance; MALDI-TOF Mass Spectrometry; Mass Spectrum Analysis; Membrane; Methods; Modeling; Pattern; Peptides; Property; Proteins; Proteomics; Range; Research; Sampling; System; Techniques; Technology; Therapeutic immunosuppression; Time; Two-Dimensional Gel Electrophoresis; UV induced; Ultrafiltration; Vacuum; animal tissue; base; capillary; cytokine; design; fibrosarcoma; human disease; in vivo; interstitial; killings; neoplastic cell; novel; secretory protein; size; tool; tumor; tumor growth

DESCRIPTION (provided by applicant): It is a challenge to obtain low abundant and/or secretory proteins/peptides (secretomes) from animals in vivo. Our laboratory has developed a novel technique to address this challenge which utilizes capillary ultrafiltration (CUF) probes to capture secretomes in vivo. Capillary ultrafiltration involves a vacuum applied to semipermeable membrane fibers to extract fluid containing secretory molecules from the interstitial space. The CUF probes also provide a simple means to obtain a small-volume sample from living biological systems. We have successfully identified and sequenced the secretomes directly by mass spectrometry, bypassing the traditional proteomics approaches which are primarily based on the 2D electrophoresis gels. In this proposal, we will use the well-established UV-induced fibrosarcoma tumor progressor/regressor model system to exploit the technique of CUF probes to collect in vivo secretomes. Tumor secretomes will be identified using mass spectrometry (MALDI-TOF MS and Q-TOF 2 MS/MS) and enzyme-linked immunosorbent assay (ELISA). In parallel, we will optimize this technique by performing high-throughput mass spectrometry (MALDI-TOF-TOF) and Cytokine Antibody Arrays. We believe that using CUF probes/Mass spectrometry will be a powerful tool which can be applied to other animal models and other human diseases.

描述（由申请人提供）： 从体内动物中获得低丰富和/或分泌蛋白/肽（分泌型）是一个挑战。我们的实验室已经开发了一种新型技术来应对这一挑战，该挑战利用毛细血管超滤（CUF）探针在体内捕获了秘密组。毛细血管超滤涉及将真空吸收到半透明的膜纤维上，以从间质空间中提取含有分泌分子的液体。 CUF探针还提供了一种简单的手段，可以从生物生物系统中获取小体积样本。我们已通过质谱直接成功地识别并测序了秘密组，绕过了主要基于2D电泳凝胶的传统蛋白质组学方法。在此提案中，我们将使用良好的紫外线诱导的纤维肉瘤肿瘤进行者/回归器模型系统来利用CUF探针的技术来收集体内秘密。将使用质谱法（MALDI-TOF MS和Q-TOF 2 ms/ms）和酶联免疫吸附测定法（ELISA）来鉴定肿瘤秘密。同时，我们将通过执行高通量质谱（MALDI-TOF-TOF）和细胞因子抗体阵列来优化此技术。我们认为，使用CUF探针/质谱法将是一种强大的工具，可以应用于其他动物模型和其他人类疾病。