PROCESSED ANTIGEN CHARACTERIZATION BY MASS SPECTROMETRY

通过质谱法进行抗原表征

• 批准号: 6373317
• 负责人: DONALD F HUNT
• 金额: $ 53.85万
• 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 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6373317
• 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：(改编自申请者摘要)继续研究 多肽序列分析仪器和方法的发展 与第I类和第I类有关的抗原呈递给免疫系统 提出了主要组织相容性复合体的II类分子。例程 疾病相关抗原的检测和表征 10,000个自体多肽混合物中的阿托莫尔水平是预期结果。 疾病相关抗原的鉴定是疾病研究的重要第一步 开发有效的疫苗或其他免疫系统调节剂 对抗细菌和病毒感染、癌症、自身免疫性疾病和组织 移植排斥反应。多阶段层析与我们的 最新开发的自动高峰停车技术，加上纳升/分钟，高 高效液相色谱和高效毛细管 通过电喷雾将电泳法与离子陷阱质谱仪对接 将使用电离源来实现这一目标。《公约》的具体目标 建议的研究包括以下内容：(1)鉴定黑色素瘤特异性 由I类分子，人类白细胞抗原A2.1，A1和A3提出的抗原，以及类 II分子，DR4，(2)，用于识别与之相关的次要组织相容性抗原 移植物抗宿主病和组织移植排斥反应，(3)识别 髓鞘碱性蛋白来源的多肽与交叉反应自身 多发性硬化症相关II类递呈的多肽 分子DR15Dw2，并被来自多个来源的T细胞克隆识别 硬化症患者，(4)鉴定I类分子呈递的多肽， B27，是强直性脊柱炎患者所特有的(5) 为了鉴定由I类分子A2.1呈现的独特的多肽 对诊断为慢性淋巴细胞白血病的个人，(6)至 鉴定和鉴定已发布的由人类白细胞抗原-A2.1呈递的多肽 通过磷酸化或糖基化进行翻译修饰，(7)到 鉴定存在的人类白细胞抗原-DR4Dw4分子呈递的II类肽 以及缺乏伴侣分子，即人类白细胞抗原-DM。