Low-volume ITC for investigating biomolecular interactions

蒙特拉尔大学的几名研究人员正在研究重要的大分子复合体，并共同组成了由NSERC资助的CREATE培训计划，名为“大分子复合体的细胞动力学”(CDMC)。这一CREATE计划旨在培养高素质的人才，以便在包括制药和生物技术行业在内的多个就业部门就业。正在研究的大分子复合体在调节生物过程中非常重要，如转录、RNA加工、糖酵解、病毒感染、抗生素耐药性、膜离子传输和癌症。为了充分了解这些重要的大分子复合体中特定相互作用的作用，彻底表征络合物形成的结合亲和力和特异性是至关重要的。在过去的几年里，在测量大分子络合物的结合亲和力方面取得了许多重大的技术进步。然而，等温滴定热法(ITC)是一种允许直接测量真实结合亲和力的技术，因为它不需要对感兴趣的分子进行任何修改，并且测量可以在溶液中进行。ITC使用标准微量热计进行实验的主要缺点是，当测量弱结合作用时，需要大量的样品材料，这往往是令人望而却步的。因此，最近开发了低容量的国贸中心仪器。低容量的ITC仪器不仅大大减少了所需的样本量，而且还增加了样品吞吐量。这些小体积的微量热计对于研究相互作用是必不可少的，当结合较弱时，当样品材料难以获得或昂贵时。作为CREATE-CDMC计划的一部分，所要求的低容量微量热计将有助于满足记录ITC大分子络合物实验以及培训学生和博士后研究员的迫切需求。

Several researchers at the Université de Montréal are investigating important macromolecular complexes and together have formed an NSERC-funded CREATE training program entitled "Cellular dynamics of macromolecular complexes" (CDMC). This CREATE program is designed to train highly qualified personnel for employment in a number of job sectors, including pharmaceutical and biotech industries. The macromolecular complexes being investigated are important in regulating biological processes such as transcription, RNA processing, glycolysis, viral infection, antibiotic resistance, ion-transport through membranes and cancer. In order to fully understand the role of specific interactions within these important macromolecular complexes, it is crucial to thoroughly characterize both the binding affinity and the specificity of complex formation. Over the last several years, there have been a number of significant technical advancements in measuring the binding affinities of macromolecular complexes. However, isothermal titration calorimetry (ITC) is one technique that allows direct measurement of true binding affinity since it does not require any modification of the molecules of interest and the measurements can be done in solution. The main disadvantage of ITC experiments with standard microcalorimeters is that, when measuring weak binding interactions, large quantities of sample material are needed, and this can often be prohibitive. As a result, low-volume ITC instruments have recently been developed. Low-volume ITC instruments not only significantly minimize the amount of sample required, but also increase sample throughput. These low-volume microcalorimeters are essential for studying interactions when the binding is weak and when sample material is either difficult to obtain or expensive. The requested low-volume microcalorimeter will help fulfill a pressing need for recording ITC experiments of macromolecular complexes and for training students and postdoctoral fellows as part of the CREATE-CDMC program.