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

尽管发现了1400多种潜在的癌症生物标志物，但由于缺乏经过良好验证的定量方法来测定蛋白质水平，将这些发现转化为临床应用的速度非常缓慢。将低丰度蛋白质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SISCAPA-MS，是一种很有前途的方法，可用于平行定量分析多种肽分析物。这种方法需要针对这些潜在生物标志物的大量蛋白型肽的高亲和力抗体的可用性。为了克服多肽抗原小鼠杂杂瘤技术的局限性，以及兔多克隆抗体的不可再生性，我们提出将兔对多肽抗原的强大免疫反应与AvantGen专有的酵母展示系统相结合，分离出高亲和力的兔抗癌相关多肽单克隆抗体。在这项II期研究中，将从酵母中分离出针对100个多肽的抗体，展示抗体文库，这些抗体文库是由从免疫过多肽偶联物的兔的脾脏或B细胞中分离出的抗体基因构建的。分离得到的抗体具有高亲和力、可再生性和生产成本低等特点。

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.