Proteomics-based Direct Diagnosis

这项NSERC合作和研究发展资助提案代表了加拿大生物技术公司温哥华岛抗体有限公司（专门从事基于SISCAPA®免疫质谱的诊断技术）与位于维多利亚大学的加拿大大学研究团队（Cameron实验室）之间令人兴奋的合作机会，该团队在螺旋体生物学和诊断抗原发现和开发方面具有新颖的专业知识。以及比利时安特卫普热带医学研究所（凯尼恩实验室）的一个研究小组，他们在蛋白质组学和生物标志物发现方面具有专长。该建议侧重于尿液和血浆样本中微生物生物标志物的测量，使用新技术，以实现活动性感染的直接诊断。由于Cameron和Kenyon实验室在螺旋体研究领域的综合专业知识，我们将重点关注四种经济上重要且全球流行的感染：梅毒（由梅毒螺旋体亚种引起），雅司病（由梅毒螺旋体亚种引起），钩端螺旋体病（由钩端螺旋体引起）和莱姆病（由伯氏疏螺旋体引起）。与其他诊断方法不同，SISCAPA®技术测量来自感染生物体的抗原，从而改善了目前不合格的诊断。该方法包括三个主要步骤：(1)鉴定在感染期间以足够水平表达的独特螺旋体蛋白，随后选择关键的蛋白型肽作为蛋白质替代品，并衍生针对这些肽的亲和纯化抗体；(2)检测抗体富集替代肽的能力，并通过质谱法分析其检测下限；(3)检测样品（尿液和血浆），以诊断活动性感染。该项目将通过培训高技能工人、加强研究和开发以及建立一个能够成功诊断具有全球分布、具有经济相关性和历史上难以诊断的疾病的诊断平台，为加拿大带来重大的经济效益。

This NSERC Collaborative and Research Development Grant proposal represents an exciting collaborative opportunity between a Canadian biotechnology company, Vancouver Island Antibodies Ltd., that specializes in SISCAPA® immuno-mass spectrometry-based diagnostic technology, a Canadian university-based research team (Cameron laboratory) located at the University of Victoria with novel expertise in spirochete biology and diagnostic antigen discovery and development, and a research team at the Institute of Tropical Medicine in Antwerp, Belgium (Kenyon laboratory) with expertise in proteomics and biomarker discovery. This proposal focuses on the measurement of microbial biomarkers in urine and plasma samples, using novel technology, to enable direct diagnosis of active infections. As a result of the combined expertise of the Cameron and Kenyon labs within the field of spirochete research, we will focus upon four economically important and globally prevalent infections: syphilis (caused by Treponema pallidum subspecies pallidum), yaws (caused by Treponema pallidum subspecies pertenue), leptospirosis (caused by Leptospira species) and Lyme disease (caused by Borrelia burgdorferi). Unlike other diagnostic methods, SISCAPA®technology measures antigens from the infecting organism thus improving on current sub-standard diagnostics. The method involves three main steps: (1) identification of unique spirochete proteins that are expressed at a sufficient levels during infection, subsequent selection of key proteotypic peptides as protein surrogates and derivation of affinity-purified antibodies specific for these peptides; (2) testing of the antibodies for their ability to enrich the surrogate peptides and analysis of their lower limits of detection by mass spectrometry and (3) testing of samples (urine and plasma) for diagnostic identification of active infection. The project will have significant economic benefit for Canada through training of highly skilled workers, enhancement of research and development and the establishment of a diagnostic platform that can successfully diagnose diseases that have a global distribution, are economically relevant and have historically been difficult to diagnose.