IDENTIFICATION OF GRAVES DISEASE SPECIFIC ANTIGENS

格雷夫斯病特异性抗原的鉴定

• 批准号: 3488332
• 负责人: DONALD W BOWDEN
• 金额: $ 5万
• 依托单位: COLLABORATIVE RESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-05-01 至 1985-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3488332
• 关键词: Escherichia coli; Graves disease; autoantibody; autoantigens; autoimmune disorder; biotechnology; complementary DNA; gene expression; genetic library; genetic manipulation; human tissue; immunochemistry; immunoelectrophoresis; immunologic assay /test; immunological substance; immunopathology chemotherapy; molecular cloning; nucleic acid sequence; radioimmunoassay

We propose to develop a methodology by which the genes coding for autoimmune disease-specific protein antigens can be rapidly cloned using recombinant DNA techniques. The system which we are using lends itself to high level expression of these antigens as hybrid proteins in E. coli. This facilitates production of these antigens for generating antibodies or developing immunodiagnostic tests. In Phase I a cDNA library will be constructed from human thyroid tissue. Cloning will be into the lambda gt11 system. Clones carrying Graves' specific sequences will be identified by differential immuno-screening with sera from Graves' patients and sera from normal individuals. Graves' positive clones can be induced with the lambda gt11 system, leading to expression of substantial amounts of the Graves' antigen fused to beta-galactosidase (hybrid protein). Because of their large size, they can subsequently be purified easily. In Phase II the cloned Graves' antigens will be characterized. They will be tested extensively for the ability to bind specifically with autoantibodies present in the sera from Graves' patients. Cloned antigens determined to be Graves' specific will be used to develop diagnostic tests directed for use in clinical diagnostic laboratories.

我们建议开发一种方法，通过基因编码