IGF::OT::IGF RECOMBINANT RABBIT MONOCLONAL ANTIBODIES AS AFFINITY CAPTURES FOR PEPTIDES

IGF::OT::IGF 重组兔单克隆抗体作为肽的亲和捕获

• 批准号: 9149418
• 负责人: XIAMON FAN
• 金额: $ 100万
• 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: 2015-09-22 至 2017-09-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9149418
• 关键词: Affinity; Antibodies; Antibody Affinity; Antigens; B-Lymphocytes; Biological Assay; Biological Markers; Genes; Hybridomas; Immune; Immune response; Keyhole Limpet Hemocyanin; Libraries; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Monoclonal Antibodies; Mus; Nature; Oryctolagus cuniculus; Peptides; Performance; Proteins; Recombinants; Spleen; System; Technology; Translations; Yeasts; clinical application; commercialization; phase 2 study; polyclonal antibody; stable isotope

Despite the discovery of over 1400 potential cancer biomarkers, translation of the discovery into clinical application has been very slow due to the lack of well-validated quantitative methods for assaying protein levels. Methods combining immune-enrichment of peptides derived from low abundance proteins and stable isotope dilution with mass spectrometry (MS), so-called SISCAPA-MS, are a promising approach for quantitative analysis of multiple peptide analytes in parallel. This method requires the availability of high affinity antibodies against large panels of proteotypic peptides for each of these potential biomarkers. To overcome the limitations of mouse hybridoma technology with peptide antigens, and the non-renewable nature of rabbit polyclonal antibodies, we propose to combine the robust immune response of rabbits to peptide antigens with AvantGen's proprietary yeast display system to isolate high affinity rabbit monoclonal antibodies against cancer-related peptides. During this Phase II study, antibodies against 100 peptides will isolated from yeast display antibody libraries constructed with antibody genes isolated from spleen or B cells from rabbits that have been immunized with peptide conjugates. The isolated antibodies will be of high affinity, renewable and can be produced in an economical manner.

尽管发现了 1400 多种潜在的癌症生物标志物，但由于缺乏用于测定蛋白质水平的经过充分验证的定量方法，将这一发现转化为临床应用的速度非常缓慢。将低丰度蛋白衍生肽的免疫富集和稳定同位素稀释与质谱 (MS) 相结合的方法（即所谓的 SISCAPA-MS）是并行定量分析多种肽分析物的一种有前途的方法。 该方法需要针对这些潜在生物标志物中的每一个的大量蛋白质肽提供高亲和力抗体。为了克服肽抗原小鼠杂交瘤技术的局限性以及兔多克隆抗体的不可再生性，我们建议将兔对肽抗原的强大免疫反应与 AvantGen 专有的酵母展示系统相结合，以分离针对癌症相关肽的高亲和力兔单克隆抗体。在这项 II 期研究中，将从酵母展示抗体库中分离出针对 100 种肽的抗体，该库是用从已用肽缀合物免疫的兔子的脾脏或 B 细胞中分离出的抗体基因构建的。分离的抗体将具有高亲和力、可再生并且可以以经济的方式生产。