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.

对纯化抗原的需求，无论是脂多糖 (LPS) 还是蛋白质，通常是 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) 被设计为一个用户指导的设施，用于 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。具有制备容易发生的蛋白质的专业知识 由于聚合性质或疏水性质而产生的聚集将基于 从志贺氏菌中生产蛋白质（在某些情况下还包括脂多糖）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， APCL 主任和联合主任将研究目标蛋白质的动态和功能特性 也极大地促进了个体研究人员所需的细菌抗原的生产。 每种待制备的蛋白质在其制备的任何给定步骤中都可能存在其自身的一系列困难， 克服这些问题已成为APCL成员的强项。 APCL未来5年的具体目标是： 1：提供克隆、质粒服务 按照 Pi 的要求构建和过度表达基因产物以用于该计划 各种项目。 2：开发和实施针对特定情况进行独特优化的定制协议 纯化用于拟议研究的过度表达蛋白质。 3：表征 表达和纯化的蛋白质在纯度、结构元素和溶液中的分散度方面如下 正确折叠的初步指标。 4：根据要求提供体细胞抗原（LPS）以供使用 研究者们在他们的个人研究中。 该核心内提供的专业知识和服务对于拟议的各个方面都是基础 研究和 APCL 的便利性将协同增强所提出的成果 提供由生产专家研究人员制作的一致质量的抗原库 此类大分子。