CCHI Antigen Purification Core

CCHI 抗原纯化核心

• 批准号: 7701570
• 负责人: WILLIAM D. PICKING
• 金额: $ 23.81万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-18 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7701570
• 关键词: Animal Model; Animals; Antigens; Attention; Bacterial Antigens; Biochemical; Burkholderia; Center for Translational Science Activities; Cloning; Custom; Development; Diagnostic; Diagnostic Procedure; Disease; Elements; Ensure; Filament; Gastrointestinal tract structure; Gene Targeting; Generations; Goals; Growth; Hand; Health; Helicobacter pylori; Human; Immune response; Immunology; Individual; Integral Membrane Protein; Investigation; Laboratories; Lead; Lipopolysaccharides; Maintenance; Measures; Mission; Mucosal Immunity; Mucous Membrane; Outcome; Physiological; Plasmids; Preparation; Prevention; Procedures; Process; Production; Property; Protein Biosynthesis; Proteins; Protocols documentation; Pseudomonas; Quality Control; Recombinant Proteins; Reproduction; Research; Research Personnel; Salmonella; Salmonella typhi; Scheme; Serotyping; Services; Shigella; Shigella dysenteriae; Solubility; Solutions; Source; Specificity; Staging; Structure; System; Therapeutic Intervention; Time; Training; Vaccines; Work; Yersinia; base; biodefense; design; driving force; enteric pathogen; experience; expression cloning; innovation; interest; macromolecule; member; microorganism; pathogen; polymerization; programs; protein function; wasting

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.

纯化抗原的需求，无论是脂多糖（LPS）还是蛋白质，通常都是障碍 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）被设计为用户指导的设施 大分子的产生可用于研究肠道病原体。提供的服务将包括 自定义蛋白质合成和LP的制备。蛋白质容易发生的专业知识 由于聚合特性或疏水性能引起的聚集 从志贺氏菌中生产蛋白质（和LP）的丰富经验， 沙门氏菌，耶尔森氏菌，假单胞菌和Burkholderia系统。对结构的理解 APCL主管和联合导演对目标蛋白的动态和功能特性将 还极大地促进了生产各个研究人员要求的细菌抗原的努力。 每种要准备的蛋白质都可以在准备的任何给定步骤中呈现自己的一组困难， 克服此类问题已成为APCL成员的力量。 未来5年中APCL的具体目的是：1：在克隆，质粒中提供服务 根据PI要求在程序中使用的基因产品的构建和过表达 各种项目。 2：为唯一优化的定制协议开发和实施 在拟议的研究中使用过表达的蛋白质的纯化。 3：表征 在纯度，结构元素和溶液中的分散性方面表达和纯化的蛋白质作为 适当折叠的初步指标。 4：根据要求提供躯体抗原（LPS） 研究人员的个人研究。 该核心内提供的专业知识和服务至关重要 研究和APCL的便利性将协同增强拟议的结果 提供一致的抗原，由调查人员创造的抗原，这些抗原是生产专家 这种大分子。