PROCESSED ANTIGEN CHARACTERIZATION BY MASS SPECTROMETRY

通过质谱法进行抗原表征

• 批准号: 6534032
• 负责人: DONALD F HUNT
• 金额: $ 48.38万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6534032
• 关键词: HY antigen; MHC class I antigen; MHC class II antigen; antigen presentation; bioimaging /biomedical imaging; biomedical equipment development; capillary electrophoresis; cross immunity; electrospray ionization mass spectrometry; graft versus host disease; high performance liquid chromatography; histocompatibility antigens; melanoma; multiple sclerosis; myelin basic proteins; protein sequence; tumor antigens

DESCRIPTION: (adapted from applicant's abstract) Research to continue the development of instrumentation and methods for the sequence analysis of peptide antigens presented to the immune system in association with class I and class II molecules of the major histocompatibility complex is proposed. Routine detection and characterization of disease associated antigens present at the attomole level in a mixture of 10,000 self peptides is the expected outcome. Identification of disease associated antigens is an important first step in the development of vaccines or other immune system modulators that are effective against bacterial and viral infections, cancer, autoimmune disorders and tissue transplant rejection. Multistage chromatography in conjunction with our recently developed, automated peak parking technology plus nanoliter/min, high performance liquid chromatography and high performance capillary electrophoresis interfaced to an ion trap mass spectrometer via an electrospray ionization source will be employed to achieve this goal. Specific aims of the proposed research include the following: (1) to identify melanoma specific antigens presented by the class I molecules, HLA A2.1, A1 and A3, and the class II molecule, DR4, (2) to identify minor histocompatibility antigens associated with graft-vs-host disease and tissue transplant rejection, (3) to identify peptides derived from myelin basic protein as well as cross reactive self peptides that are presented by the multiple sclerosis associated class II molecule, DR15Dw2, and recognized by T-cell clones derived from multiple sclerosis patients, (4) to identify peptides presented by the class I molecule, B27, that are unique to individuals diagnosed with ankylosing spondylitis (5) to identify peptides presented by the class I molecule, A2.1, that are unique to individuals diagnosed as having chronic lymphocytic leukemia, (6) to identify and characterize peptides presented by HLA-A2.1, that have been post translationally modified by either phosphorylation or glycosylation, (7) to identify class II peptides presented by the HLA-DR4Dw4 molecule in the presence and absence of the chaperone molecule, HLA-DM.

描述：（改编自申请人的摘要）研究继续 仪器的开发和肽序列分析的方法 与I类和类联合的免疫系统呈现的抗原 提出了主要的组织相容性复合物的II分子。常规 疾病相关抗原的检测和表征 预期的结果是10,000个自肽的混合物中的attomole水平。 鉴定疾病相关抗原是重要的第一步 开发有效的疫苗或其他免疫系统调节剂 针对细菌和病毒感染，癌症，自身免疫性疾病和组织 移植排斥。多阶段色谱与我们的结合 最近开发的，自动化的峰值停车技术加上纳米尺/分钟，高 性能液相色谱和高性能毛细管 通过电喷雾连接到离子陷阱质谱仪的电泳 电离源将用于实现这一目标。特定目标 拟议的研究包括以下内容：（1）鉴定特异性黑色素瘤 由I类分子呈现的抗原HLA A2.1，A1和A3以及类 II分子，DR4，（2），以识别相关的小组织相容性抗原 使用移植物vs宿主疾病和组织移植排斥，（3）以识别 源自髓磷脂碱性蛋白以及交叉反应性自我的肽 由多发性硬化症相关的II类提出的肽 分子，DR15DW2，并由T-Cell克隆识别。 硬化症患者（4）确定I类分子提出的肽， B27，这是诊断为强直性脊柱炎的个体所独有的（5） 识别I级分子A2.1所呈现的肽，它们是独特的 对于被诊断为慢性淋巴细胞性白血病的个体，（6）至 识别和表征由HLA-A2.1提出的肽， 通过磷酸化或糖基化的翻译修饰，（7）至 在存在下识别由HLA-DR4DW4分子提出的II类肽 伴侣分子的缺失HLA-DM。