CCHI Antigen Purification Core

对纯化抗原的需求，无论是脂多糖(LPS)还是蛋白质，往往是阻碍 有效开发针对细菌病原体的诊断方法或预防措施。 抗原生产核心实验室(APCL)被设计为用户指导的设施，用于 在肠道病原体研究中有用的大分子的产生。提供的服务将包括 定制蛋白质合成和脂多糖的制备。在制备容易感染的蛋白质方面的专业知识 由于聚合性质或疏水性质而引起的聚集将基于 在从志贺氏菌中生产蛋白质(在某些情况下还有内毒素)方面有丰富的经验， 沙门氏菌、耶尔森氏菌、假单胞菌和伯克霍尔德氏菌系统。对结构的理解， APCL主任和联席主任将介绍目标蛋白的动态和功能特性 还极大地便利了生产个别调查人员所要求的细菌抗原的努力。 每种要制备的蛋白质在其制备过程中的任何给定步骤都会出现其自身的一组困难， 克服这些问题已经成为APCL成员的一项优势。 APCL在未来5年的具体目标是：1：提供克隆、质粒化服务 根据PI‘s的要求构建和过度表达用于计划的基因产品 各种各样的项目。2：开发和实施定制协议，专门针对 提纯过表达的蛋白质以用于拟议的研究。3：特色化 表达和纯化蛋白质的纯度、结构元素和在溶液中的分散性 折叠正常的初步指标。4：应要求提供体细胞抗原(LPS)，供 调查人员在他们的个人研究中。 该核心内提供的专业知识和服务是建议的所有方面的基础 APCL的研究和便利性将通过以下方式协同增强拟议成果 提供由生产专家的研究人员制造的一致质量的抗原池 这样的大分子。

The need for purified antigen, whether lipopolysaccharide (LPS) or protein, is often a roadblock for the efficient development of diagnostic methods or preventative measures targeting bacterial pathogens. The Antigen Production Core Laboratory (APCL) is designed as a user-directed facility for the generation of macromolecules useful in the study of enteric pathogens. Services rendered will include custom protein synthesis and the preparation of LPS. Expertise with the preparation of proteins prone to aggregation due to polymerizing properties or hydrophobic properties will be provided based upon extensive experience with the production of proteins (and LPS in some cases) from the Shigella, Salmonella, Yersinia, Pseudomonas and Burkholderia systems. An understanding of the structural, dynamic, and functional properties of targeted proteins by the Director and Co-Director of the APCL will also greatly facilitate efforts in producing the bacterial antigens requested by the individual investigators. Each protein to be prepared can present its own set of difficulties at any given step in its preparation, and overcoming such problems has become a strength of the members of the APCL. The Specific Aims for the APCL over the next 5 years are: 1: To provide services in cloning, plasmid construction and over-expression of gene products as requested by the Pi's for use in the Program's various projects. 2: To develop and implement customized protocols uniquely optimized for the purification of the over-expressed proteins to be used in proposed studies. 3: To characterize expressed and purified proteins in terms of purity, elements of structure, and dispersity in solution as preliminary indicators of proper folding. 4: To provide somatic antigen (LPS) as requested for use by the investigators in their individual studies. The expertise and services provided within this Core are fundamental to all aspects of the proposed research and the convenience of the APCL will synergistically enhance the proposed outcomes by providing a consistent quality pool of antigens made by investigators who are experts in the production of such macromolecules.