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: 7694362
• 负责人: John Charles Chaput
• 金额: $ 16.39万
• 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 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7694362
• 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; Peptides; Positioning Attribute; Process; Production; Prostate-Specific Antigen; Protein Analysis; Proteins; Proteome; 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（由申请人提供）：监测大量蛋白质水平的能力为疾病指标（生物标志物）提供了检测和治疗许多疾病（包括癌症）的巨大希望。尽管目前用于检测人血清中蛋白质水平的大多数分析技术依赖于基于ELISA的测定或质谱分析，但蛋白质捕获阵列作为用于蛋白质组范围分析的未来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（合成体），其可用于检测和分析人血液和唾液中的癌症和癌症相关蛋白。与免疫球蛋白相比，其高成本和低生产速率限制了合适质量亲和试剂的可用性，从低成本化学试剂快速合成人工抗体为蛋白质配体问题提供了可能的解决方案。该申请的目标是使用我们称之为配体和接头系统重组（SRLL）的技术开发针对五种已知癌症生物标志物的合成抗体。该技术将表面等离子体共振（SPR）的多重能力与DNA自组装的纳米级精度相结合，以鉴定从先前通过筛选或选择方法鉴定的单价配体产生多价结合剂所需的最佳肽对和肽对分离距离。我们将从该过程中出现的配体称为“合成体”。该提案的具体目标是： 1.确定产生对五种公认的癌症生物标志物具有中等亲和力的肽配体所需的最少选择步骤数。 2.针对特定目标1中靶向的五种癌症生物标志物生成高亲和力合成抗体阵列。拟议研究的相关性在于基于常规蛋白质组分析的早期癌症检测的潜力，使用合成抗体阵列在症状前检测癌症生物标志物。

项目成果

Creating protein affinity reagents by combining peptide ligands on synthetic DNA scaffolds.

通过在合成DNA支架上结合肽配体来产生蛋白质亲和力试剂。

• DOI: 10.1021/ja9051735
• 发表时间: 2009-12-02
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Williams, Berea A. R.;Diehnelt, Chris W.;Belcher, Paul;Greving, Matthew;Woodbury, Neal W.;Johnston, Stephen A.;Chaput, John C.
• 通讯作者: Chaput, John C.

Synthesis of peptide-oligonucleotide conjugates using a heterobifunctional crosslinker.

• DOI: 10.1002/0471142700.nc0441s42
• 发表时间: 2010-09
• 期刊: Current protocols in nucleic acid chemistry
• 作者: Williams, Berea A R;Chaput, John C
• 通讯作者: Chaput, John C

Generating DNA synbodies from previously discovered peptides.

• DOI: 10.1002/cbic.201100284
• 发表时间: 2011-08-16
• 期刊: CHEMBIOCHEM
• 影响因子: 3.2
• 作者: Liu, Rui;Jiang, Bing;Yu, Hanyang;Chaput, John C.
• 通讯作者: Chaput, John C.