Synthesis of MUC1 glycopeptides and generation of antibodies for the indentificat

描述（由申请人提供）：MUC 1糖肽的合成和用于鉴定癌细胞上表位的抗体的产生。大多数癌过度表达细胞表面糖蛋白，这些糖蛋白被截短的O-聚糖异常糖基化，这是肿瘤相关抗原。健康细胞和癌细胞的细胞表面组成的这种明显差异使得肿瘤相关抗原成为开发癌症特异性免疫疗法的有吸引力的靶标。近年来发现糖肽结合表位是肿瘤免疫治疗的理想靶点。在所有异常糖基化的癌细胞表面糖蛋白中，研究得最好的是粘蛋白“MUC 1”，然而，由癌细胞动态表达的免疫原性MUC 1糖肽的结构特征仍然研究不足。分析表位的糖部分和肽部分的工具是不可用的。为了解决这个问题，本文提出的研究结合了候选糖肽的化学合成（含有糖肽的14个Tn或STn的文库），以及单克隆抗体的产生，这些抗体随后将用于特异性识别癌细胞表面表位（长期目标）。此外，这些探针将用于研究随着癌症进展表位表达的变化。该SC2试验项目的目的是合成MUC 1糖肽文库，以产生第一组单克隆抗体探针，并验证它们与它们所引发的糖肽的结合特异性。一套完整的单克隆抗体的可用性将对开发新的癌症特异性免疫疗法、诊断和疫苗产生重大影响。所有糖肽将在P.I.中合成。在埃尔帕索的德克萨斯大学的实验室。私家侦探的实验室还将在埃尔帕索得克萨斯大学的边界生物医学核心设施确定单克隆抗体对MUC 1衍生糖肽的结合亲和力和结合特异性。将与Dapeng Zhou，M.D.，哲学博士、黑色素瘤和内科肿瘤学系;德克萨斯大学医学博士安德森癌症中心免疫学系。 公共卫生相关性：MUC 1是一种细胞表面糖蛋白，在许多癌症中过度表达和异常糖基化。存在于细胞表面上的肿瘤相关抗原是开发有效免疫疗法的理想靶标，然而，癌细胞表达的表位的确切结构特征尚不清楚。我们的目标是开发一套能够特异性识别这些细胞表面糖肽表位的单克隆抗体。

DESCRIPTION (provided by applicant): Synthesis of MUC1 glycopeptides and generation of antibodies for the identification of epitopes on cancer cells. Most carcinomas over-express cell surface glycoproteins that are aberrantly glycosylated with truncated O-glycans, which are tumor-associated antigens. This distinct difference in cell surface make-up of healthy and cancerous cells make tumor- associated antigens attractive targets for the development of cancer-specific immunotherapies. Recently, it was discovered that combined sugar and peptide epitopes make excellent targets for immunotherapy of cancer. Of all aberrantly glycosylated cancer cell surface glycoproteins the one best studied is the mucin "MUC1", however, the structural features of immunogenic MUC1 glycopeptides, dynamically expressed by cancer cells, remain understudied. Tools to analyze the sugar part and peptide part of an epitope are unavailable. In order to address this question the research proposed here combines the chemical synthesis of candidate glycopeptides (a library of 14 Tn or STn containing glycopeptides), and the generation of monoclonal antibodies, which will later be used to specifically identify cancer cell surface epitopes (long-term goal). Furthermore, these probes will be used to study changes in epitope expression as the cancer progresses. The objective of this SC2 pilot project is to synthesize a MUC1 glycopeptide library, to generate a first set of monoclonal antibody probes, and to validate their binding specificity to the glycopeptides they have been elicited against. The availability of a comprehensive set of such monoclonal antibodies would have major implications for the development of novel cancer-specific immunotherapies, diagnostics and vaccines. All glycopeptides will be synthesized in the P.I.'s laboratory at the University of Texas at El Paso. The P.I.'s laboratory will also determine the binding affinities and binding specificities of the monoclonal antibodies to the MUC1 derived glycopeptides at the Border Biomedical Core Facility at the University of Texas at El Paso. Mice will be immunized and monoclonal antibodies will be raised in collaboration with Dapeng Zhou, M.D., Ph.D., Department of Melanoma and Medical Oncology; Department of Immunology, University of Texas MD Anderson Cancer Center in Houston, TX. PUBLIC HEALTH RELEVANCE: MUC1 is a cell surface glycoprotein that is over-expressed and aberrantly glycosylated in many carcinoma. The tumor-associated antigens present on the cell surface are ideal targets for the development of effective immunotherapies, however, the exact structural features of the epitopes expressed by cancer cells are not known. It is our goal to develop the tools, a set of monoclonal antibodies, which can specifically identify these cell surface glycopeptide epitopes.