A High-throughput Route to Synthetic Antibodies for Array-based Cancer Detection

用于基于阵列的癌症检测的合成抗体的高通量途径

• 批准号: 7384541
• 负责人: John Charles Chaput
• 金额: $ 19.85万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-29 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7384541
• 关键词: Affinity; Antibodies; Binding; Biological Assay; Biological Markers; Blood; CA-125 Antigen; Cancer Detection; Carcinoembryonic Antigen; Chemicals; Classification; DNA; Detection; Disease; Enzyme-Linked Immunosorbent Assay; Escherichia coli Proteins; Figs - dietary; Future; Genetic Recombination; Goals; Human; Individual; Length; Libraries; Ligands; Malignant Neoplasms; Messenger RNA; Methods; Mind; Monitor; Numbers; Peptides; Positioning Attribute; Process; Production; Prostate-Specific Antigen; Protein Analysis; Proteins; Proteome; Rate; Reagent; Research; Route; Saliva; Screening for cancer; Screening procedure; Serum; Solid; Solutions; Specificity; Surface; Surface Plasmon Resonance; Techniques; Technology; alpha-Fetoproteins; base; cost; interest; nanoscale; novel strategies; numb protein; protein expression; prototype; scaffold; self assembly

DESCRIPTION (provided by applicant): The ability to monitor the levels of large numbers of proteins for indicators of disease (biomarkers) holds great promise for the detection and treatment of many diseases, including cancer. Although most analytical techniques for detecting protein levels in human serum currently rely on ELISA-based assays or mass spectroscopic analysis, protein capture arrays have gained considerable interest as a future technology for proteome-wide analysis. The studies described in this application seek to develop a comprehensive array of high affinity synthetic antibodies (synbodies) that can be used to detect and profile cancer and cancer-related proteins in human blood and saliva. In contrast to immunoglobins, for which high costs and slow production rates limit the availability of suitable quality affinity reagents, the rapid synthesis of artificial antibodies from low cost chemical reagents offers a possible solution to the protein ligand problem. The goal of this application is to develop synthetic antibodies to five known cancer biomarkers using a technology that we call systematic recombination of ligands and linkers (SRLL). This technology combines the multiplex capability of surface plasmon resonance (SPR) with the nanometer-scale precision of DNA self-assembly to identify optimal peptide pairs and peptide pair separation distances needed to produce multivalent binding agents from monovalent ligands previously identified by screening or selection methods. We have termed the ligands that emerge from this process "synbodies". The Specific Aims of this proposal are to: 1. Determine the minimum number of selection steps needed to produce modest affinity peptide ligands to five well-established cancer biomarkers. 2. Generate an array of high affinity synthetic antibodies to the five cancer biomarkers targeted in Specific Aim 1.The relevance of the proposed research resides in the potential for early cancer detection based on routine proteome-wide analysis using synthetic antibody arrays to detect cancer biomarkers presymptomatically.

描述(申请人提供)：监测疾病指示物(生物标记物)的大量蛋白质水平的能力对包括癌症在内的许多疾病的检测和治疗具有很大的前景。尽管目前大多数检测人血清中蛋白质水平的分析技术依赖于基于ELISA法的分析或质谱分析，但蛋白质捕获阵列作为一种未来的蛋白质组分析技术已经获得了相当大的兴趣。本申请中描述的研究寻求开发一系列全面的高亲和力合成抗体(综合体)，可用于检测和分析人类血液和唾液中的癌症和癌症相关蛋白。免疫球蛋白的高成本和缓慢的生产速度限制了合适的高质量亲和试剂的可获得性，而从低成本的化学试剂快速合成人工抗体为蛋白质配体问题提供了一种可能的解决方案。这项应用的目标是利用一种我们称为配体和连接物的系统重组(SRLL)的技术来开发针对五个已知癌症生物标记物的合成抗体。这项技术结合了表面等离子体共振(SPR)的多重能力和DNA自组装的纳米级精度，可以从先前通过筛选或选择方法确定的单价配体中识别出生产多价结合剂所需的最佳多肽对和多价多肽对分离距离。我们把这个过程中产生的配体称为“复合体”。这项建议的具体目的是： 1.确定产生与五个公认的癌症生物标记物的中等亲和力的多肽配体所需的最少选择步骤。 2.产生针对特定目标的五个癌症生物标记物的高亲和力合成抗体阵列1.拟议的研究的相关性在于基于常规的蛋白质组分析的早期癌症检测的可能性，该分析使用合成抗体阵列以症状前检测癌症生物标记物。