High Throughput Immunoassay for Flagellin

鞭毛蛋白的高通量免疫分析

• 批准号: 8516169
• 负责人: MARTIN D. CHAPMAN
• 金额: $ 18.43万
• 依托单位: INDOOR BIOTECHNOLOGIES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8516169
• 关键词: Academia; Accounting; Adjuvant; Admission activity; Affect; Affinity; Allergens; Allergic; Allergic Disease; Animals; Antibodies; Asthma; Atopic Dermatitis; Bacteria; Bacterial Proteins; Biological Assay; Biological Markers; Biotechnology; Canis familiaris; Centers for Disease Control and Prevention (U.S.); Child; Childhood; Chronic Disease; Collaborations; Communicable Diseases; Complement; Detection; Development; Dictyoptera; Disease; Dose; Dust; Endotoxins; Environmental Exposure; Environmental Monitoring; Enzyme-Linked Immunosorbent Assay; Europe; Exclusion; Exposure to; Family; Farming environment; Felis catus; Flagella; Flagellin; Genetic; Goals; Government Agencies; Hay fever; High Pressure Liquid Chromatography; Home environment; Hospitalization; Hospitals; House Dust; Household; Housing; Human; Hypersensitivity; Immune response; Immunoassay; Immunoblotting; Immunology; Infectious Disease Immunology; Investigation; Ion Exchange; Life; Link; Low Prevalence; Manufacturer Name; Marketing; Measurement; Measures; Molds; Molecular Weight; Monoclonal Antibodies; Mus; National Institute of Environmental Health Sciences; Natural Immunity; Oryctolagus cuniculus; Patients; Performance; Pharmacologic Substance; Phase; Precipitation; Production; Proteins; Pseudomonas aeruginosa; Public Health; Reagent; Recombinants; Reproducibility; Research; Research Personnel; Risk Factors; Role; Sampling; Sensitivity and Specificity; Services; Site; Small Business Innovation Research Grant; Socioeconomic Status; Sodium Chloride; Standardization; Suspension substance; Suspensions; System; Techniques; Testing; Toll-Like Receptor 5; Ultrafiltration; Vaccines; air sampling; allergic response; base; commercialization; cooking; early childhood; economic cost; environmental allergen; mouse model; polyclonal antibody; protective effect; public health relevance; pyroglyphid; research study; respiratory; response; tool

DESCRIPTION (provided by applicant): Asthma is a serious public health problem in the US that affects one in ten schoolchildren and causes ~500,000 hospitalizations each year, with an estimated economic cost of $56 billion. Allergic sensitization to household allergens (dust mites, cat, dog, mouse, cockroach, molds) is a major risk factor for the development of asthma. Asthma in early childhood can also be influenced by exposure to bacterial products which may potentiate or have protective effects depending on the level of environmental exposure and genetic factors. Recent studies in a mouse model have shown that the bacterial protein, flagellin, promotes allergic sensitization to indoor allergens and that human patients with asthma have high levels of antibodies to flagellin. Household dust samples have been shown to contain flagellin. However, investigation of the association between exposure to flagellin and asthma requires a robust test to measure flagellin in environmental samples from homes and public buildings. The goal of this SBIR Phase I project is to develop a high throughput immunoassay for flagellin, capable of analyzing several hundred samples per week. The specific aims of the project are i) to purify flagellin from Pseudomonas aeruginosa; ii) to develop a monoclonal or polyclonal antibody based immunoassay which detects flagellin from most bacterial species; iii) to validate the assay performance to measure flagellin in dust samples of known allergen and endotoxin content. The strategy involves developing a multiplexed immunoassay which can be used to measure flagellin, allergens and molds simultaneously. This assay will be to assess the role of flagellin exposure in allergic disease. Commercial potential: Successful completion of this SBIR will result in valuable immunoassay product(s) that can be marketed by the applicant, Indoor Biotechnologies, to allergy/immunology researchers in academia, government agencies, pharmaceutical companies and vaccine manufacturers, worldwide. The research could generate several product lines that are complementary to the company's current products and which will enable the company to expand its product range in new markets.

描述（由申请人提供）：哮喘是美国严重的公共卫生问题，影响十分之一的学龄儿童，每年导致约50万人住院，估计经济成本为560亿美元。对家庭过敏原（尘螨、猫、狗、小鼠、蟑螂、霉菌）的过敏性致敏是哮喘发展的主要风险因素。儿童早期哮喘也可能受到暴露于细菌产物的影响，这些细菌产物可能会增强或具有保护作用，这取决于环境暴露和遗传因素的水平。最近在小鼠模型中的研究表明，细菌蛋白鞭毛蛋白促进对室内过敏原的过敏性致敏，并且患有哮喘的人类患者具有高水平的鞭毛蛋白抗体。家庭灰尘样本已被证明含有鞭毛蛋白。然而，研究暴露于鞭毛蛋白和哮喘之间的关联需要一个强大的测试来测量家庭和公共建筑的环境样品中的鞭毛蛋白。SBIR第一阶段项目的目标是开发一种高通量的鞭毛蛋白免疫测定方法，能够每周分析数百个样品。该项目的具体目标是：i）从铜绿假单胞菌中纯化鞭毛蛋白; ii）开发一种基于单克隆或多克隆抗体的免疫测定法，可检测大多数细菌物种的鞭毛蛋白; iii）验证测定已知过敏原和内毒素含量的粉尘样品中鞭毛蛋白的测定性能。该策略涉及开发一种多重免疫测定法，可用于同时测量鞭毛蛋白、过敏原和霉菌。本试验将评估鞭毛蛋白暴露在过敏性疾病中的作用。商业潜力：成功完成此 SBIR将产生有价值的免疫测定产品，可由申请人Indoor Biotechnologies销售给全球学术界、政府机构、制药公司和疫苗制造商的过敏/免疫学研究人员。这项研究可能会产生几个产品线，这些产品线是对公司现有产品的补充，并将使公司能够在新市场中扩大其产品范围。