Rapid scanning stopped flow spectrometer circular dichroism capability tool and gel electrophoresis-transblot system

我们阿尔伯塔大学土木工程与环境工程系的研究小组在现有的快速扫描停止流(RS-SF)分光光度计的基础上添加了圆二色谱(CD)功能以及凝胶电泳-转印系统，这将是非常有用和迫切需要的。Prion(即蛋白质感染性颗粒)是一组神经退行性疾病的病原体，称为可传递海绵状脑病(TSE)，如牛海绵状脑病(BSE)。自2005年1月在加拿大出生的奶牛中首次出现疯牛病以来，加拿大养牛业一直在生产大量特定的风险材料(SRM)，以将疯牛病对人类健康、食品安全和动物健康造成的风险降至最低。由于Pron蛋白在环境中的潜在持久性和独特的稳定性，迫切需要开发有效的去污方法来去除简单和复杂基质中的Pron蛋白。目前正在测试臭氧和高级氧化处理方法来破坏感染性绵羊瘙痒病Pron蛋白以及Pron模型蛋白。推测臭氧化和AOPS可以成功地破坏类病毒蛋白的一级和二级结构。这一假设可以通过使用现有的RS-SF分光光度计和将要购买的新的CD模块的CD光谱以及使用新的凝胶电泳-转印系统的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹分析以及我们实验室现有的液体色谱-离子陷阱串联质谱仪来验证。这个Pron灭活研究计划最终将导致开发出非常有效和经济可行的臭氧和高级氧化处理工艺来处理SRM处置的废物，并将有助于保护加拿大的野生动物、动物饲养操作和公众健康。

A circular dichroism (CD) capability added to an existing rapid scanning stopped flow (RS-SF) spectrophotometer along with a gel electrophoresis-transblot system, will be highly useful and urgently needed for our research group in the Department of Civil & Environmental Engineering at the University of Alberta to conduct research on prion inactivation.Prions (i.e., proteinaceous infectious particles) are causative agents of a group of neurodegenerative diseases called transmissible spongiform encephalopathies (TSEs) such as bovine spongiform encephalopathy (BSE) in cattle. Since the first occurrence of BSE in a Canadian-born cow in January 2005, the cattle industry in Canada has been producing large amounts of specific risk materials (SRMs) to minimize the risks that BSE causes to human health, food safety and animal health. Because of potential persistence and unique stability of prion proteins in the environment, there is an urgent need for developing effective decontamination methods for prion protein in simple and complex matrices.Ozonation and advanced oxidation treatments are being tested for the destruction of infectious sheep scrapie prion, as well as prion-model proteins. It is speculated that the primary and secondary structures of prion-like proteins can be successfully disrupted by ozonation and AOPs. This hypothesis can be tested by CD spectroscopy using an existing RS-SF spectrophotometer coupled with a new CD module to be purchased and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis using the new gel electrophoresis-transblot system along with an existing liquid chromatography-ion trap tandem mass spectrometer in our laboratory.This prion inactivation research program will eventually lead to a development of very effective and economically feasible ozone and advanced oxidation treatment processes for the wastes from SRMs disposal, and will help protect Canadian wildlife, animal feeding operations, and public health.