Developing a highly specific and sensitive glucagon ELISA kit

开发高度特异性和敏感性的胰高血糖素 ELISA 试剂盒

• 批准号: 8974749
• 负责人: XIAOMIN FAN
• 金额: $ 22.47万
• 依托单位: AVANTGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8974749
• 关键词: Address; Affect; Affinity; Amino Acids; Antibodies; Antibody Affinity; Antigen Targeting; Antigens; B-Lymphocytes; Binding; Biological Assay; Biology; Blood; Blood Circulation; Blood Glucose; C-terminal; Cells; Cleaved cell; Custom; Cysteine; DNA amplification; Detection; Diabetes Mellitus; Ensure; Enteroglucagon; Enzyme-Linked Immunosorbent Assay; Epidemic; Exhibits; GLP-2; Glucagon; Gluconeogenesis; Glucose; Glycolysis; Goals; Harvest; Hormones; Human; Hypoglycemia; Immunization; Immunoglobulin Variable Region; Individual; Injection of therapeutic agent; Insulin; Keyhole Limpet Hemocyanin; Kinetics; Label; Libraries; Light; Liver; Measurement; Measures; Methods; Monitor; Monoclonal Antibodies; Mus; N-terminal; OGTT; Oryctolagus cuniculus; Pancreas; Pathway interactions; Peptides; Peripheral Blood Mononuclear Cell; Plasma; Play; Prediabetes syndrome; Preparation; Process; Production; Protocols documentation; RNA; Reagent; Relative (related person); Reverse Transcriptase Polymerase Chain Reaction; Role; Specificity; Spleen; Splenocyte; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Tissues; Yeasts; blood glucose regulation; carboxyl radical; cross reactivity; glucagon-like peptide 1; novel; peptide hormone; phase 1 study; proglucagon; prototype; public health relevance; response; therapeutic target; vector

 DESCRIPTION (provided by applicant): With prediabetes and diabetes reaching epidemic proportions, there is an urgent need to identify more effective therapeutics. Diabetes is characterized by elevated levels of the peptide hormone glucagon,, and inappropriate suppression of glucagon during an oral glucose tolerance test. Glucagon plays a major role in up-regulating glucose release from the liver during hypoglycemic conditions, and serves as a key counter-regulatory hormone to insulin. Consequently, glucagon pathways are an attractive target for therapeutic intervention. However, tissue-specific processing of the proglucagon precursor results in the production of several other peptide hormones, such as oxyntomodulin, GLP-1 and GLP- 2, that also affect glucose levels but by different direct or indirect modes of action. The presence of several alternatively cleaved products that also harbor the glucagon sequence along with the low levels (< 50pM) of glucagon in the circulation has hampered efforts to develop a suitable assay for accurate determination of glucagon levels when analyzing potential therapeutic targets. The goal of this proposal is to use AvantGen's novel yeast display system and rabbit monoclonal antibody discovery technology to isolate pM affinity, highly specific anti-glucagon antibody clones to be used to develop a highly specific and sensitive assay reagents.

 描述(申请人提供)：随着糖尿病前期和糖尿病达到流行程度，迫切需要确定更有效的治疗方法。糖尿病的特征是多肽激素胰高血糖素水平升高，以及在口服葡萄糖耐量试验中不适当地抑制胰高血糖素。在低血糖状态下，胰高血糖素在上调肝脏释放葡萄糖方面起着重要作用，并且是胰岛素的关键逆调节激素。因此，胰升糖素途径是治疗干预的一个有吸引力的靶点。然而，对原胰高血糖素前体的组织特异性处理会导致其他几种多肽激素的产生，如氧合调节蛋白、GLP-1和GLP-2，这些激素也会影响血糖水平，但作用方式不同。几种选择性裂解产物的存在也含有胰高血糖素序列，以及循环中低水平(50pM)的胰高血糖素，这阻碍了在分析潜在治疗靶点时开发一种准确测定胰高血糖素水平的合适方法的努力。本方案的目标是利用AvantGen的新型酵母展示系统和兔单抗发现技术来分离PM亲和力强、高度特异的抗胰高血糖素抗体克隆，以用于开发一种高度特异和敏感的检测试剂。