CCHI Antigen Purification Core

对纯化抗原的需要，无论是脂多糖（LPS）还是蛋白质，通常是免疫治疗的障碍。 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）设计为用户指导的设施， 产生可用于研究肠道病原体的大分子。提供的服务将包括 定制蛋白质合成和LPS的制备。蛋白质制备的专业知识， 由于聚合性质或疏水性质的聚集将基于 具有从志贺氏菌生产蛋白质（和某些情况下的LPS）的丰富经验， 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， 由APCL的Director和Co-Director对靶蛋白的动态和功能特性进行分析， 也极大地促进了个体研究者所需的细菌抗原的生产。 每种待制备的蛋白质在其制备中的任何给定步骤都可能存在其自身的一组困难， 克服这些问题已成为APCL成员的强项。 未来5年，APCL的具体目标是：1.提供克隆，质粒， 基因产物的构建和过表达，如PI所要求的，用于该计划的 各种项目。2：开发和实施定制的协议， 纯化待用于所提出的研究的过表达蛋白。3：定性 表达和纯化的蛋白质的纯度，结构元素，和分散性在溶液中， 正确折叠的初步指标。4：根据要求提供体细胞抗原（LPS），供 研究人员在他们的个人研究。 本核心内提供的专门知识和服务对拟议的 研究和APCL的便利性将协同提高拟议的成果， 提供由生产方面的专家研究者制备的质量一致的抗原池 这些大分子的。

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.