Characterization of Innate Binding Specificities of Peptidoglycan Binding Protein

肽聚糖结合蛋白的固有结合特异性的表征

• 批准号: 8882250
• 负责人: Andrew E. Levin
• 金额: $ 21.13万
• 依托单位: IMMUNETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8882250
• 关键词: Antibodies; Applications Grants; Autoimmune Diseases; Bacillus cereus; Bacteria; Bacterial Infections; Basic Science; Binding; Binding Proteins; Binding Sites; Biological Assay; Blood Platelets; Buffers; Cell Wall; Cells; Chemistry; Cleaved cell; Clinical Research; Color; Complementary DNA; Densitometry; Detection; Diagnosis; Disease; Drosophila pros protein; Employee Strikes; Enterococcus faecalis; Enzymes; Escherichia coli; Excision; Goals; Gram-Negative Bacteria; Gram-Positive Bacteria; HIV; Health; Hemocytes; Hemolymph; High Pressure Liquid Chromatography; Human; Immune system; Inflammation; Insecta; Klebsiella pneumonia bacterium; Larva; Length; Lepidoptera; Lysostaphin; Malignant Neoplasms; Manduca; Manduca sexta; Measures; Membrane; Membrane Proteins; Meningitis; Methods; Microbe; Monophenol Monooxygenase; Muramic Acid; Muramidase; Mutate; Natural Immunity; Nosocomial Infections; Oklahoma; Oryctolagus cuniculus; Pathogen detection; Peptidoglycan; Performance; Polyacrylamide Gel Electrophoresis; Polysaccharides; Precipitation; Protein Binding; Proteins; Quality Control; Reaction; Recombinant Proteins; Recombinants; Research; Risk; Role; Sampling; Sepsis; Septic Shock; Serine Protease; Source; Specificity; Staphylococcus aureus; Supplementation; Surface; Technology; Testing; Thick; Transfusion; Universities; Vertebral column; base; blood product; complement system; crosslink; improved; innovation; microbial; mutanolysin; pathogen; peptidoglycan recognition protein; polyclonal antibody; pro-phenoloxidase; protein expression; research study; screening; systems research

DESCRIPTION (provided by applicant): BacTx(R) technology, developed by Immunetics for detection of bacteria, employs a unique and innovative assay chemistry to detect Gram-positive and Gram-negative bacteria. The technology is based on components of the innate immune system of the tobacco hornworm (Manduca sexta), in particular the prophenoloxidase cascade, in which serine proteases cleave prophenoloxidase to phenoloxidase, which subsequently oxidizes chromogen to generate a colored reaction product. The BacTx(R) Bacterial Detection Kit received 510(k) clearance from the FDA (CBER) in June 2012 as a quality control test for bacterial contamination in platelets. For M. sexta and other Lepidoptera, the most potent elicitor of the prophenoloxidase cascade is peptidoglycan, which is a major structural component of bacterial cell walls. In this application we propose to biochemically characterize the relationship between the presence of specific bacterial cell wall features and evasion of detection by pathogen recognition proteins present in M. sexta hemolymph. The role of these features in inhibiting prophenoloxidase activation will be investigated, with the long term goal of potentially enhancing assay performance and opening up new applications of BacTx(R) technology, including potential use for diagnosis of sepsis, meningitis, and nosocomial infections. Due to the striking similarities between the insect innate immune system and human complement system, this research may also prove significant to elucidating the relationship between inflammation due to bacterial infection and debilitating disorders such as cancer, autoimmune diseases, and septic shock.

描述（由申请人提供）：BacTx(R)技术是由Immunetics公司开发的用于检测细菌的技术，采用独特和创新的分析化学方法来检测革兰氏阳性和革兰氏阴性细菌。该技术基于烟草角虫（Manduca sexta）先天免疫系统的组成部分，特别是酚氧化酶原级联，其中丝氨酸蛋白酶将酚氧化酶原裂解为酚氧化酶，酚氧化酶随后氧化色素生成有色反应产物。BacTx(R)细菌检测试剂盒于2012年6月获得FDA （CBER）的510(k)许可，作为血小板细菌污染的质量控制测试。对于sexta和其他鳞翅目来说，最有效的酚氧化酶原级联的激发子是肽聚糖，它是细菌细胞壁的主要结构成分。在这个应用中，我们建议用生物化学方法来描述这种关系