High Performance Liquid Chromatography system

这是一个申请购买自动进样器高效液相色谱(HPLC)系统。高效液相色谱仪可以从复杂的蛋白质或多肽混合物中非常有效地(高分辨率)分离和分离蛋白质和多肽。我实验室的这项研究(由NSERC资助)的一个方面是描述与细胞分化过程(精子发生)有关的蛋白质，这些蛋白质导致男性精子细胞(精子)的形成。我们还对卵子受精后这些蛋白质的特性和识别机制感兴趣。这项工作需要使用高效液相色谱对不同脊椎动物和无脊椎动物的蛋白质进行纯化和表征，这对不孕症具有重要的潜在意义。这些蛋白质的进一步表征是通过在特定的蛋白酶的帮助下将它们裂解成更小的多肽来实现的。生成的多肽混合物也可以方便地用高效液相色谱法进行分级。这样得到的纯肽被用于鉴定目的蛋白。这一信息非常有价值，因为它可以进一步用于编码这些蛋白质的基因的表征。高效液相色谱法是几乎所有致力于蛋白质纯化的现代生化实验室中常规使用的技术。在过去的23年里，我们的实验室每天都在使用这项技术。然而，非常出人意料的是，在我们实验室运行高效液相色谱法的主板最近死了，需要更换它的部件已经过时了。这为我们提供了获得带有自动取样器的现代系统的机会，此外还允许更换对有效实施我们当前的研究计划至关重要的设备。自动取样器将大大提高该仪器的多用户可用性，这将有利于UVic生物化学和微生物学系的其他几位同事进行的免疫学(关于昏睡病的研究)和微生物学(钩端螺旋体病/痘病毒)的研究。因此，利用该设备进行的研究，除了其强大的基础研究组成部分外，可能对加拿大和国际上的人们具有重要的应用有益影响。

This is an application to purchase autosampler High Performance Liquid Chromatography (HPLC) system. HPLC allows a very efficient (high resolution) fractionation and isolation of proteins and peptides from complex protein or peptide mixtures. One aspect of the research (NSERC funded) in my lab is to characterize the proteins that are involved in the cell differentiation process (spermatogenesis) leading to the formation of male sperm cells (spermatozoa). We are also interested in the characterization and to identify the mechanisms of these proteins after fertilization of the egg. This work requires the purification and characterization of proteins from different vertebrate and invertebrate organisms using HPLC and has important potential implications for infertility. Further characterization of these proteins is achieved by cleaving them into smaller peptides with the help of specific proteases. The peptide mixtures generated can also be conveniently fractionated by HPLC. The pure peptides thus obtained are used in the identification of the proteins of interest. This information is very valuable as it can be further used in the characterization of the genes encoding these proteins. HPLC is a technique that is routinely used in almost all modern biochemistry labs working on protein purification. Our lab has been using this technique on a daily basis for the last twenty three years. However, quite unexpectedly the mother board that runs HPLC in our lab died recently and the parts required to replace it have become obsolete. This provides us the opportunity to acquire a modern system with an autosampler in addition to allowing for the replacement of an equipment that is imperative for the efficient implementation of our current research program. The autosampler will greatly enhance the multiuser availability of this instrumentation which will benefit the immunology (studies on sleeping sickness) and microbiology (leptospirosis/poxvirus) research conducted by several other colleagues at the Department of Biochemistry and Microbiology at UVic. Hence, the research pursued with this equipment, besides its strong basic research component, may have important applied beneficial implications for people in Canada and Internationally.