CORE FACILITY RESEARCH PEPTIDE SYNTHESIZER

细胞和分子生物学中的许多重要进展可归因于蛋白质的结构-功能分析，所述蛋白质控制单个细胞或细胞群体中的功能。固相肽合成（SPPS）的小肽，或确定的结构，功能，或较大的蛋白质的免疫原性结构域，使表征这些分子内的结构-活性关系（SAR），否则不能实现。Harbor-UCLA研究和教育研究所的一系列研究人员越来越依赖肽合成，这是目前努力获得组合SPPS仪器的基础，以扩大Harbor-UCLA REI生物聚合物合成核心设施的能力并使其现代化。作为该核心设施的一个组成部分，拟议的仪器适用于各种各样的肽合成。因此，由该仪器产生的肽将作为一系列生物医学研究项目的优秀试剂。例如，这样的肽对于进行SAR研究、通过流式细胞术检查肽-靶细胞相互作用和肽的原位定位是非常宝贵的。同样，SPPS能够生产含有不寻常的烷基化、α、β-氨基和D-对映体氨基酸的肽，这些氨基酸对于研究与肽或蛋白质功能相关的构象至关重要。现代表达系统和定点诱变方法对于常规和快速生产这样的肽是不可行的。所提出的仪器的组合合成能力将通过同时合成多个不同的肽来提高我们的核心设施的效率。由所提出的仪器生成的用于NIH资助项目的未标记或标记肽试剂的应用包括：1）蛋白质或肽功能结构域扫描; 2）使用非放射性13 C-和15-N-肽文库和FTIR或NMR光谱评估蛋白质和肽内的关键SAR; 3）用标记或未标记肽探测的表达文库或突变克隆的筛选; 4）设计和合成具有抗微生物、抗肿瘤或免疫增强特性的肽用于最终的治疗评价;和5）合成抗原或诊断肽试剂以检测或表征用于疫苗开发的免疫原靶。将使用一种简单的算法来简化仪器的操作和管理，该算法协调从在线肽合成请求到向研究者交付质量受控肽到再充电核算的整个过程。Harbor-UCLA REI最大限度地支持该仪器作为生物聚合物合成核心设施的一部分的长期成功。因此，拟议的工具将显着提高效率和多样性的SPPS所需的NIH赞助和假设驱动的研究。

Many important advances in cellular and molecular biology are attributable to structure-function analysis of proteins that .govern function in individual cells or populations of cells. Solid phase peptide synthesis (SPPS) of small peptides, or defined structural, functional, or immunogenic domains of larger proteins, has enabled characterization of structure-activity relationships (SAR) within these molecules that could not be achieved otherwise. The increasing reliance on peptide synthesis by an array of investigators at the Harbor-UCLA Research and Education Institute is the basis for the current effort to acquire a combinatorial SPPS instrument, to expand and modernize the capabilities of the Biopolymer Synthesis Core Facility at Harbor-UCLA REI. As an integral component of this Core Facility, the proposed instrument is adaptable to a wide diversity of peptide syntheses. Thus, peptides produced by this instrument will serve as excellent reagents for a broad array of biomedical research projects. For example, such peptides are invaluable for conducting SAR studies, examining peptide- target cell interactions by flow cytometry, and localization of peptides in situ. Likewise, SPPS enables the production of peptides containing unusual alkylated, alpha,beta-dehydro, and D-enantiomer amino acids crucial for studying conformation as it relates to peptide or protein function. Contemporary expression systems and site-directed mutagenesis approaches are not feasible for routine and rapid production of such peptides. The combinatorial synthesis capability of the proposed instrument will enhance the efficiency of our Core Facility by enabling simultaneous synthesis of multiple, distinct peptides. Applications for unlabeled or labeled peptide reagents to be generated by the proposed instrument for use in NIH-sponsored projects include: 1) protein or peptide functional domain scanning; 2) evaluation of key SAR within proteins and peptides using non-radioactive 13 C- and 15-N-peptide libraries and FTIR or NMR spectroscopy; 3) screening of expression libraries or mutant clones probed with labeled or unlabeled peptides; 4) design and synthesis of peptides with antimicrobial, antitumor, or immunopotentiating properties for eventual therapeutic evaluation; and 5) synthesis of antigen or diagnostic peptide reagents to detect or characterize immunogen targets for vaccine development. Operation and management of the instrument will be streamlined using a simple algorithm coordinating the entire process from on-line peptide synthesis request, to delivery of quality-controlled peptides to investigators, to recharge accounting. The Harbor-UCLA REI is maximally supportive of the long-term success of the instrument as part of the Biopolymer Synthesis Core Facility. Therefore, the proposed instrument will significantly enhance the efficiency and diversity of SPPS required for NIH-sponsored and hypothesis-driven research.