Multiplexed vaccine titer test

描述（由申请人提供）：到1990年，免疫接种工作几乎消除了脊髓灰质炎，麻疹和腮腺炎等儿童疾病，但新疫苗的快速引入可能会促进重新出现，因为复杂的注射时间表降低了父母和医生的依从性。疫苗制造商已经用多价疫苗做出了回应，这些疫苗提供了针对多种病原体的单次注射保护。除麻疹、腮腺炎和风疹疫苗（MMR）外，白喉类毒素、破伤风类毒素和无细胞百日咳（DTaP）、灭活脊髓灰质炎病毒（IPV）、B型流感嗜血杆菌（Hib）和/或B肝炎（HepB）的五价组合疫苗已获得批准，其他疫苗仍在开发中。疫苗的范围和多价性的增加产生了对具有成本效益的、高度多重的血清学测试的需求，以确定对每种组分的抗体应答。需要新的测试作为筛选测定来评估每个患者的保护，并作为疫苗开发的工具来检测新组合中每个靶点的功效。 传统的多孔ELISA形式的免疫测定是费力的，并且与多分析物测试的匹配性差。流式细胞术珠阵列免疫测定可以用小型化、多重化、快速、灵敏和特异性的替代方法代替这种ELISA形式的测定。尽管用于血清学的珠阵列具有监测疫苗接种和感染的商业潜力，但与多重夹心免疫测定如细胞因子组相比，该市场仍未开发。一个关键的限制是需要用不同的抗原包被珠子，包括蛋白质、多糖和全病毒颗粒，这需要广泛的固定化化学，必须独立优化。由于全面的疫苗接种和感染测定将需要>50重测试，因此制备稳健且可再现的珠板的挑战是一个重大障碍。 该提交材料提出，通过开发允许采用新型化学物质和检测设计对不同抗原进行快速多重检测的方案来克服当前检测的局限性，并扩展微珠阵列免疫测定的潜力，以包被微珠，而不受抗原是碳水化合物、蛋白质还是病毒的影响。这将导致用于筛选对所有批准的疫苗的反应和暴露于最常见的传染性生物体的试剂盒。重要的是，这些试剂盒是完全动态的，并且可以进行修改以反映新疫苗、新传染病血清型和/或新发现的疾病生物体。这个平台有可能被设计用于第三世界。 公共卫生相关性：多价疫苗滴度试验尚未开发出快速测定多价疫苗应答水平的标准化方法。标准化测试的可用性将允许更有效地开发新的多价疫苗以及确定疫苗在接种儿童和成人中的有效性。在本次提交的文件中，我们提议开发一种平台技术，该技术将允许对疫苗功效进行快速、经济有效且灵敏的标准化测试。

DESCRIPTION (provided by applicant): Immunization efforts had all but eliminated childhood diseases such as polio, measles, and mumps by 1990, but the rapid introduction of new vaccines may paradoxically be promoting reemergence as complex injection schedules decrease compliance by parents and physicians. Vaccine manufacturers have responded with multivalent vaccines that provide single-injection protection against multiple agents. In addition to the measles, mumps and rubella vaccine (MMR), pentavalent combinations of diptheria toxoid, tetanus toxoid, and acellular pertussis (DTaP), inactivated polio virus (IPV), Haemophilus influenzae Type b (Hib), and/or hepatitis B (HepB), have been approved and others remain in development. The increasing range and multivalency of vaccines has created a need for cost-effective, highly multiplexed serological testing to determine antibody response to each component. New tests are needed both as screening assays to evaluate protection of each patient and as a tool for vaccine development to detect efficacy for each target in novel combinations. The traditional multiwell ELISA format immunoassay is laborious and poorly matched to multianalyte testing. Flow cytometric bead array immunoassays can replace such ELISA format assays with a miniaturized, multiplexed, rapid, sensitive and specific alternative. Despite the commercial potential of bead arrays for serology to monitor vaccination and infection, this market remains undeveloped compared to multiplexed sandwich immunoassays such as cytokine panels. A key limitation is the need to coat beads with diverse antigens, including proteins, polysaccharides, and whole virus particles, which requires a wide range of immobilization chemistries that must be independently optimized. Insofar as a comprehensive vaccination and infection assay will require >50-plex testing, the challenge of preparing robust and reproducible bead panels is a significant barrier. This submission proposes to overcome the limitations of current assays and expand the potential of the bead array immunoassay by the development of protocols that allow rapid multiplexing of diverse antigens employing novel chemistries and assay design to coat beads independent of whether the antigen is a carbohydrate, protein or virus. This will lead to kits for screening for responses to the full range of approved vaccines and for exposure to the most common infectious organisms. Importantly, these kits are completely dynamic and can be modified to reflect new vaccines, new infectious disease serotypes and/or newly discovered disease organisms. This platform has the potential to be engineered for use in the third world. PUBLIC HEALTH RELEVANCE: Multivalent Vaccine Titer Test A standardized method to rapidly determine the level of response to multivalent vaccines has not been developed. Availability of a standardized test would allow more efficient development of new multivalent vaccines as well as determining effectiveness of vaccines in vaccinated children and adults. In this submission we propose the development of a platform technology that will allow rapid, cost effective and sensitive standardized testing for vaccine efficacy.