CORE--SOLID PHASE PEPTIDE SYNTHESIS AND HPLC/CZE

该核心将合成和表征肽，并开发新的HPLC和cz分析系统，以提高分辨率和效率。此外，该核心将执行圆二色（CD）实验的要求。使用Merrifield自动化SPPS， Core每年将合成大约20个肽（长度为10到50个残基，线性，环状，磷酸化）。采用最新的肽/蛋白制备纯化技术，我们将提供10 ~ 100mg纯化肽/蛋白进行HPLC、FPLC离子交换层析和毛细管区带电泳（CZE）。必要时将使用质谱（LSIMS）和Edman降解（Core C）进行表征。这些肽将支持大多数项目，而Core C将使用CZE。多肽将用于进行功能和结构研究，以及提高特异性多克隆抗体。最后，我们打算利用这个核心的一些资源来开发新的分析技术和探索新的应用。这个核心将利用P，I。他的经验（12%的努力）和他的技术助理的经验，以及他的功能齐全的SPPS和表征实验室。实验室空间配备通风柜和其他小型设备。主要仪器包括一台HX-110型LSI质谱仪和JEOL公司的60 MHz核磁共振仪，一台贝克曼公司的P/ACE型2050 CZE，三台贝克曼公司的肽合成器，分析和制备hplc和冻干机。私家侦探将主持用户委员会，负责设定优先级。预计Core D将能够以先到先得的方式运作。在必须确定优先级的情况下，将执行用户委员会大多数成员（主席除外）达成的决定。多肽的合成和纯化将由M. odroaz - sypniewski进行（30%的努力）。D. Kirby将负责HPLC， FPLC和CZE的质量控制和方法开发（15%的努力）。质谱分析（15%的工作）将由A. Craig博士（15%的工作）进行。在相关私人顾问的同意下，多余的肽将免费分发给任何NIH资助的研究人员，就像我们过去所做的那样。近年来，人们对圆二色光谱的需求越来越大。我们负责该仪器已有十多年，并将为该计划项目的调查人员提供分析。

This Core will synthesize and characterize peptides, and develop new HPLC- and CZE-based analytical systems to improve on resolution and efficiency. Additionally, this Core will perform circular dichroism (CD) experiments as requested. Using the Merrifield automated SPPS, Core will synthesize, each year, a total of approximately 20 peptides (10 to 50 residues in length; linear, cyclic, phosphorylated). Using the latest techniques for the preparative purification of peptide/proteins, we will provide 10 to 100 mg of purified peptides/proteins to HPLC, ion exchange chromatography on FPLC and capillary zone electrophoresis (CZE). Characterization will use mass spectroscopy (LSIMS) and Edman degradation (Core C) when necessary. These peptides will support most projects will Core C will use CZE. Peptides will be used to carry out functional and structural studies as well as to raise specific polyclonal antibodies. Finally, we intend to use some of the resources of this Core to develop new analytical techniques and explore new applications. This Core will take advantage of the P.,I.'s experience (12% effort) and that of his technical assistant as well as his fully functional laboratory for SPPS and characterization. Laboratory space equipped with hoods and other small equipment is available. Major instruments available include an LSI Mass Spectrometer Model HX-110 and a 60 MHz NMR from JEOL, a Beckman P/ACE model 2050 CZE, three peptide synthesizers from Beckman, analytical and preparative HPLCs and lyophilizers. The P.I. will chair the users' committee responsible for setting priorities. It is expected that Core D will be able to operate on a first come, first served basis. In cases where priority will have to be determined, the decision reached by the majority of the users' committee (excluding the chair) will be implemented. Synthesis and purification of peptides will be carried out by M. Odrowaz-Sypniewski (30% effort). Quality control and method development using HPLC, FPLC and CZE will be carried out by D. Kirby (15% effort). MS analyses (15% effort) will be carried out by Dr. A. Craig (15% effort). With the consent of the concerned P.I.s, excess peptides will be distributed free of charge to any NIH sponsored researcher as we have done in the past. Recently, there has been a significant need for circular dichroism spectroscopy. We have been responsible for the instrument for more than ten years and will provide analyses to investigators on this Program Project.