FPLC System for Bioactive Peptides Research

生物活性多肽的分离、纯化、功能鉴定和体内测试需要使用快速蛋白质液相色谱(FPLC)系统进行定性和定量分析。在马尼托巴大学食品和人类营养科学系，只有一个FPLC系统，该系统是在2004年购买的，但由于该型号已停产，而且马尼托巴大学校园没有合适的FPLC系统，因此不再使用。因此，这项提议的目的是用一个新的FPLC系统取代不能使用的16年前的系统，该系统将为我的NSERC资助的发现生物活性多肽研究计划的继续运行提供关键工具。FPLC提供了最快速和最具成本效益的方法来分离和分离多肽，这些多肽的形式可以表征结构-功能特性，或者在更高的压力系统(HPLC)上进一步纯化。此外，FPLC有助于快速定量估计确定的分子大小的多肽，这是结构阐明的关键组成部分，也是我的NSERC资助的Discovery研究计划的主要目标。该研究计划的重点是阐明食品蛋白衍生生物活性多肽的结构-功能特性，这些生物活性多肽通常包含在称为蛋白质水解物的食品蛋白质的酶消化中。由于酶解物含有一种(有时是数百种)多肽混合物，成功进行结构-功能分析的最初速度决定步骤是在FPLC系统上进行分离。因此，FPLC是我的探索研究工作以及成功培训高素质人员(HQP)所必需的组成部分。FPLC将被用来将生物活性多肽分离成适合于用液相色谱-串联质谱仪进行结构鉴定(氨基酸测序)的级分。对氨基酸序列的了解是发现高效多肽所必需的，这些多肽可以作为各种代谢途径的有效调节器。因此，FPLC系统是我寻求提供关于生物活性多肽作用机制的基本科学信息以及发现可以减少或防止负面代谢事件的新肽的宝贵工具。该系统还将促进我寻求发现高功能食品成分的研究，这些成分可以改善加拿大人的营养状况，同时促进加拿大农作物和动物产品的附加值利用。这是因为从仪器中获得的结果将为研究人员提供设计新的食品成分的能力，这些成分可以用于配制功能食品和营养食品。FPLC系统将由研究生和本科生、博士后研究员/研究助理和技术人员每天使用，用于多肽的分级和定量。因此，该工具对HQP的成功培训至关重要。

Isolation, purification, functional characterization, and in vivo testing of bioactive peptides require qualitative and quantitative analysis using the fast protein liquid chromatography (FPLC) system. At the Department of Food and Human Nutritional Sciences, University of Manitoba, there is only one FPLC system, which was purchased in 2004 but is no longer serviceable because the model has been discontinued and there is no suitable FPLC system on the University of Manitoba campuses. Therefore, the aim of this proposal is to replace the non-serviceable 16-year old system with a new FPLC system that will provide critical tools for continued operation of my NSERC-funded Discovery research program on bioactive peptides. The FPLC provides the most rapid and cost-effective means of separating and isolating peptides in forms that can be characterized for structure-function properties or further purified on a higher pressure system (HPLC). Moreover, the FPLC facilitates rapid quantitative estimation of peptides of defined molecular sizes, which is a critical component of structural elucidation and a major goal of my NSERC-funded Discovery research program. The research program focuses on elucidation of the structure-function properties of food protein-derived bioactive peptides, which are usually contained in enzymatic digests of food proteins called protein hydrolysates. Because enzymatic hydrolysates contain a mixture (sometimes hundreds) of peptides, the initial rate-determining step for successful structure-function analysis is separation on an FPLC system. Thus, the FPLC is a required component of my Discovery research work and for successful training of highly qualified personnel (HQP). The FPLC will be used to separate bioactive peptides into fractions that are suitable for structural elucidation (amino acid sequencing) by liquid chromatography-tandem mass spectrometry. Knowledge of the amino acid sequence is required for the discovery of highly potent peptides that can serve as effective modulators of various metabolic pathways. Thus the FPLC system is a valuable tool in my quest to provide fundamental scientific information on the mechanism of action of bioactive peptides as well as the discovery of novel peptides that can reduce or prevent negative metabolic events. The system will also facilitate my research that seeks discovery of highly functional food ingredients that can enhance nutritional status of Canadians while promoting value-added utilization of Canadian crops and animal products. This is because results obtained from the instrument will provide researchers with the ability to design novel food ingredients that can be used to formulate functional foods and nutraceuticals. The FPLC system will be used by graduate and undergraduate students, postdoctoral fellows/research associates, and technicians on a daily basis for peptide fractionation and quantification. Therefore, the instrument is critical to the successful training of HQPs.