Glycoconjugate based diagnostics for bacterial toxins

基于糖缀合物的细菌毒素诊断

• 批准号: 7678970
• 负责人: Alison A. Weiss
• 金额: $ 39.3万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7678970
• 关键词: Affect; Affinity; Animal Model; Animals; Antibodies; Antibody Affinity; Bacterial Toxins; Binding; Binding Sites; Biological Assay; Biotin; Blood; Botulinum Toxins; Carbohydrates; Cause of Death; Cells; Chemistry; Clostridium botulinum; Clostridium perfringens; Clostridium perfringens epsilon toxin; Complex; Complex Mixtures; Coupled; Detection; Development; Devices; Diagnostic; Disease Outbreaks; Effectiveness; Electronics; Elements; Feces; Generations; Glycoconjugates; Goals; Gold; High temperature of physical object; Human; Individual; Intervention; Lead; Libraries; Life; Ligands; Link; Medical; Outcome; Patients; Police; Property; Public Health; Research; Research Personnel; Ricin; Saliva; Sampling; Sensitivity and Specificity; Shiga Toxin; Site; Specimen; Speed; Staphylococcal Enterotoxin B; Streptavidin; Structure; Technology; Testing; Therapeutic; Time; Toxic effect; Toxin; Training; Treatment Efficacy; Urine; Variant; Work; aerosolized; analog; base; biothreat; botulinum; cost effectiveness; design; drug candidate; emergency service responder; falls; improved; innovation; mouse model; point of care; point-of-care diagnostics; pre-clinical; prevent; programs; prophylactic; receptor; research clinical testing; response; sensor; small molecule; tissue culture; user-friendly

DESCRIPTION (provided by applicant): The Biothreat toxins include the most lethal agents ever described. Nanogram amounts of aerosolized or injected toxins cause death. To prevent loss of life, detection of biothreat toxin must be made rapidly, accurately and with exquisite sensitivity. In addition, an ideal diagnostic should be stable and simple enough for individuals with limited medical training, such as first responders (police and fire-fighters) to use on site. Current "gold standard" detection assays are time consuming and sample dependent, and fall far short of the ideal. Our preliminary studies have shown that glycoconjugate ligands mimicking the natural toxin receptor can be used for rapid and precise detection of closely related toxins. As proof of principle, we have developed tailored carbohydrates that can discriminate between the two antigenic forms of Shiga toxin, Stx1 and Stx2. We propose to extend these studies to develop diagnostic and therapeutic products. In Aim 1, we will develop 2nd and 3rd generation toxin ligands specific for Shiga, Ricin, Botulinum, Clostridium perfingens epsilon toxins and Staphylococcus enterotoxin B (SEB). These ligands will maximize cooperative binding and optimize differentiation between variants in each toxin class. In Aim 2 we will develop and evaluate different diagnostic platforms. Biotin will be covalently linked to the ligands. Conjugation to prefabricated streptavidin coated microtiter plates yields a glycoconjugate microarray cartridge, ideal for rapid testing of toxins in a point-of-care setting. In Aim 3, we will validate and perform pre-clinical evaluation of lead compounds from Aim 1. We will quantify the effectiveness of the microarray diagnostics and their potential as therapeutics in tissue culture, in mouse models, and in human specimens ex vivo. Sensitivity, specificity, rapidity, ruggedness, ease-of-use and cost-effectiveness will be evaluated to establish proof of principle.

描述（由申请人提供）：生物威胁毒素包括迄今为止所描述的最致命的物质。纳克量的雾化或注射毒素会导致死亡。为了防止生命损失，生物威胁毒素的检测必须快速、准确且具有极高的灵敏度。此外，理想的诊断应该足够稳定和简单，可供接受过有限医疗培训的个人（例如现场急救人员（警察和消防员））使用。目前的“金标准”检测分析非常耗时且依赖于样本，并且远远达不到理想的水平。我们的初步研究表明，模仿天然毒素受体的糖复合物配体可用于快速、精确地检测密切相关的毒素。作为原理证明，我们开发了定制的碳水化合物，可以区分志贺毒素的两种抗原形式 Stx1 和 Stx2。我们建议扩展这些研究以开发诊断和治疗产品。在目标 1 中，我们将开发针对志贺毒素、蓖麻毒素、肉毒杆菌、产气荚膜梭菌 epsilon 毒​​素和葡萄球菌肠毒素 B (SEB) 特异性的第二代和第三代毒素配体。这些配体将最大化协同结合并优化每个毒素类别中变体之间的差异。在目标 2 中，我们将开发和评估不同的诊断平台。生物素将与配体共价连接。与预制的链霉亲和素包被的微量滴定板缀合产生复合糖微阵列盒，非常适合在护理点环境中快速检测毒素。在目标 3 中，我们将对目标 1 中的先导化合物进行验证和临床前评估。我们将量化微阵列诊断的有效性及其在组织培养、小鼠模型和离体人体样本中作为治疗药物的潜力。将评估灵敏度、特异性、快速性、耐用性、易用性和成本效益，以建立原理证明。