High Throughput Immunoassay for Flagellin

描述(由申请人提供)：哮喘是美国一个严重的公共卫生问题，每十个学龄儿童中就有一个受到影响，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、老鼠、蟑螂、霉菌)的过敏性致敏是哮喘发生的主要危险因素。儿童早期哮喘也可能受到接触细菌制品的影响，细菌制品可能会增强或具有保护作用，具体取决于环境暴露水平和遗传因素。最近在小鼠模型中的研究表明，细菌蛋白质鞭毛蛋白促进了对室内过敏原的过敏反应，人类哮喘患者对鞭毛蛋白的抗体水平很高。家庭灰尘样本已被证明含有鞭毛蛋白。然而，要研究鞭毛蛋白暴露与哮喘之间的关系，需要进行强有力的测试，以测量来自家庭和公共建筑的环境样本中的鞭毛蛋白。这个SBIR第一阶段项目的目标是开发一种高通量鞭毛蛋白免疫分析方法，能够每周分析数百个样本。该项目的具体目标是：i)从铜绿假单胞菌中提纯鞭毛蛋白；ii)开发一种基于单抗或多克隆抗体的免疫分析方法，用于检测大多数细菌物种中的鞭毛蛋白；iii)验证检测已知过敏原和内毒素含量的粉尘样本中鞭毛蛋白的检测性能。该战略涉及开发一种多重免疫分析方法，可用于同时检测鞭毛蛋白、过敏原和霉菌。这项测试将评估鞭毛暴露在变态反应性疾病中的作用。商业潜力：成功完成这一任务 SBIR将产生有价值的免疫分析产品(S)，该产品可由申请者室内生物技术公司向世界各地的学术界、政府机构、制药公司和疫苗制造商的过敏/免疫学研究人员销售。这项研究可能会产生几个产品线，这些产品线是对该公司现有产品的补充，将使该公司能够在新市场扩大其产品范围。

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.