IMPROVING THE EFFICIENCY OF SAMPLE HANDLING AND PROCESSING FOR HT PDS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该项目旨在为HT PDS提供样品制备和处理方面的必要支持。样品制备的第一阶段是蛋白质表达、纯化和自旋标记，目前至少在我们合作者的一些实验室中以高效率完成。也就是说，每周有几十种突变是可行的。在ACERT，需要根据需要在当地储存和处理样品，例如，旋转标记和新鲜复溶至脂质体中。在这方面，我们还将开发与装载程序兼容并满足批量样品处理一致性要求的适当样品容器。 随着样品流量的预期大幅增加和快速周转的要求，我们将需要增加我们的样品储存和处理能力。此外，并非所有合作者都能够有效地自旋标记其蛋白质，因此这些任务在ACERT解决，因为我们拥有专业知识，并且可以使用CW和脉冲ESR准确方便地测定自旋标记效率。因此，该项目的成功在很大程度上取决于子项目0269中详述的适当设施的开发。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is to provide essential support for HT PDS in terms of sample preparation and handling. The first stage of sample preparation is protein expression, purification, and spin labeling, which is currently accomplished in at least some of the labs of our collaborators with the high efficiency. That is several dozen mutations per week are feasible. At ACERT the samples need to be stored and processed locally as needed, e.g. spin-labeled and freshly reconstituted into liposomes. We shall also develop in this context adequate sample containers compatible with the loading procedures and satisfying the consistency requirements of batch sample processing. With the expected large increase in sample flow and the requirements of fast turn-around, we shall need to increase our sample storage and handling capacities. Also, not all collaborators are able to spin-label their proteins or do it efficiently, therefore those tasks are addressed at ACERT, since we have the expertise and also can assay spin labeling efficiency accurately and expediently using CW and pulse ESR. The success of this project thus heavily relies on the development of suitable facility detailed in subproject 0269.