Novel Therapeutic Metabolite Screening Method

新型治疗代谢物筛选方法

基本信息

批准号：
6401858
负责人：
YEVGENYA AKSELBAND
金额：
$ 16.19万
依托单位：
ONE CELL SYSTEMS, INC
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-08-01 至 2002-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6401858
关键词：
Escherichia coli aquatic organism biotherapeutic agent drug discovery /isolation drug screening /evaluation flow cytometry high throughput technology saltwater environment technology /technique development

Escherichia coli aquatic organism biotherapeutic agent drug discovery /isolation drug screening /evaluation flow cytometry high throughput technology saltwater environment technology /technique development

项目摘要

An increasing number of marine microorganisms are being studied as a potential source of novel metabolites with unique biotherapeutic activity. Current methods involve isolating marine microorganisms from environmental samples, obtaining cell extracts or culture supernatants, using cellular or biochemical assays to analyze the extracts or supernatants for secreted metabolites, and isolating high secretors by serial dilutions. These methods are time consuming and labor intensive. In Phase I research, single cell gel microdrop (GMD) encapsulation technology will be combined with a unique receptor-ligand screening format and fluorescence activated cell sorting (FACS) to permit rapid isolation of individual marine microorganisms based on secretory activity, which is currently not possible. Using FACS, sub-populations of secretors as small as 1 percent can be rapidly isolated. Phase I research will develop the GMD assay using endotoxin-secreting E. coli and validate the assay using well characterized marine microorganism isolates which secrete endotoxin. Phase I research will also evaluate use of high-speed sorters to increase screening throughput. The GMD assay will help to streamline the search for potential new drug candidates derived from natural marine environments. PROPOSED COMMERCIAL APPLICATION: The proposed assay will facilitate drug discovery by permitting direct isolation of marine microorganisms based on secretory activity.

正在研究越来越多的海洋微生物作为潜在来源具有独特的生物治疗活性的新型代谢产物。当前方法涉及隔离来自环境样品的海洋微生物，获取细胞提取物或培养物上清液，使用细胞或生化测定法分析提取物或分泌代谢物的上清液，并通过连续隔离高秘书稀释。这些方法是耗时和劳动量的。在第一阶段研究，单细胞凝胶微旋转（GMD）封装技术将是与独特的受体配体筛选格式和荧光结合激活的细胞分选（FACS）允许快速隔离单个海洋基于分泌活动的微生物，目前是不可能的。使用FACS，可以迅速的1％的秘密分组的子组成孤立。第一阶段研究将使用内毒素分泌开发GMD分析大肠杆菌并使用良好特征的海洋微生物验证测定法分泌内毒素的分离物。第一阶段研究还将评估使用高速分类器以增加筛选吞吐量。 GMD分析将有助于简化寻找自然派生的潜在新药候选者的搜索海洋环境。拟议的商业应用：拟议的测定法将通过直接隔离来促进药物发现基于分泌活动的海洋微生物。