Development of Immunological Reagents for the Identification of New World Monkey Biomarkers

项目摘要 本R24 OD 030215行政补充申请寻求资金， Cytek北方光光谱流式细胞仪。所要求的仪器配备有三个 激发激光线，每个激光器16个检测器，自动样品加载器和基本启动 培训套件;仪器将存放在科学与创新中心（CSI） 位于德克萨斯州圣安东尼奥的Trinity大学。“新世界猴子”的目标是 免疫试剂资源”父奖是为了提高新的翻译价值， 世界猴（NWM）物种绒猴（MAR），松鼠猴（SQM）和猫头鹰猴 (OLM)通过产生新的单克隆抗体和能够鉴定重要的 炎症和代谢的生物标志物，缺乏试剂。需要 该工具基于以下原因：1）它将显着改善 开发在母公司裁决的具体目标2中提出的免疫测定法; 这些流式细胞术测定应用于十种选定靶中的六种（IL-4，IL-6，IL-10，IP-10， 颗粒酶B和CD 69），并将扩大由于使用 陈旧的仪器2)它将通过以下方式提高这些测定的精确性和再现性： 将所有三种NWM物质的检测开发集中在一个实验室中;管理 我们的一个合作机构的变化减少了获得更现代仪器的机会， 将检测限制在仅一个物种。3)它将从有限的科学产出中产生更深层次的科学产出， 可以从这些小猴子中回收的细胞数量;光谱流式细胞术允许 使用与传统流式细胞术不相容的常见荧光染料， 可以使用存档的光谱数据，这减少了用于控制管的细胞数量。4)它 将为培养本科生和技术人员开辟新的途径， 对光谱流式细胞术的实际理解，这是一项革命性的技术， 免疫分型; Trinity大学这种仪器的存在将使人们获得新的 高年级生物学学生的技术。光谱流式细胞仪的能力， 在单个细胞中检测大量标记物将提高识别多个细胞的能力 在小的血液或组织样品中的亚群，这对于提高 这些小身体的NWM物种。为长期维护提供机构支持 这些设备。

PROJECT SUMMARY This R24 OD030215 Administrative Supplement request seeks funds for the acquisition of a Cytek Northern Lights spectral flow cytometer. The requested instrument is equipped with three excitation laser lines, 16 detectors per laser, an automatic sample loader, and basic start-up Training Kit; the instrument will be housed in the Center for Sciences and Innovation (CSI) building of Trinity University in San Antonio, Texas. The goal of the “New World Monkey Immunoreagent Resource” parent award is to improve the translational value of the New World Monkey (NWM) species marmoset (MAR), squirrel monkey (SQM), and owl monkey (OLM) by generating novel monoclonal antibodies and assays able to identify important biomarkers of inflammation and metabolism, for which there is scarcity of reagents. The need for this instrument is based on the following reasons: 1) It will significantly improve the development of the immunological assays proposed in Specific Aim 2 of the parent award; these flow cytometry assays apply to six of the ten selected targets (IL-4, IL-6, IL-10, IP-10, granzyme B, and CD69) and will expand the limited assay scope caused by the use of antiquated instrumentation. 2) It will increase the rigor and reproducibility of these assays by concentrating assay development for all three NWM species in one laboratory; administrative changes in one of our collaborating institutions reduces access to more modern instruments and limits assays to just one species. 3) It will generate deeper scientific output from the limited number of cells that can be recovered from these small monkeys; spectral flow cytometry allows the use of common fluorescent dyes that are incompatible in conventional flow cytometry and can use archived spectral data, which reduces the number of cells used for control tubes. 4) It will generate new avenues for training undergraduate students and technical personnel in the practical understanding of spectral flow cytometry, a technique that has revolutionized immunophenotyping; presence of this instrument at Trinity University will give access to novel technologies to upper division Biology students. The capacity of the spectral flow cytometer to detect a large number of markers in a single cell will increase the ability to identify multiple cell subsets in small blood or tissue samples, which is critical for improving the translational value of these small body NWM species. Institutional support is provided for the long-term maintenance of this equipment.