Characterization of antitumor auto-antibodies using combinatorial peptide librari

使用组合肽文库表征抗肿瘤自身抗体

• 批准号: 8454976
• 负责人: WLODEK MANDECKI
• 金额: $ 27.24万
• 依托单位: PHARMASEQ, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-25 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8454976
• 关键词: Acetonitriles; Adsorption; Advanced Malignant Neoplasm; Affect; Affinity; Amino Acid Sequence; Amino Acids; Antibodies; Autoantibodies; Binding; Biological; Biological Markers; Buffers; Cancer Patient; Cancerous; Chemicals; Classification; Clinical; Collaborations; Complementarity Determining Regions; Complex; Development; Diagnostic; Diagnostic Neoplasm Staging; Discrimination; Disease; Early Diagnosis; Electromagnetics; Fingerprint; Fluorescence; Frequencies; Goals; Human; Immunoglobulins; Individual; Laboratories; Libraries; Ligands; Light; Malignant Neoplasms; Malignant neoplasm of ovary; Methods; Molecular Profiling; Organic solvent product; Outcome; Pattern; Peptide Library; Peptide Synthesis; Peptides; Phase; Positioning Attribute; Process; Proteins; Proteome; Radio; Randomized; Reader; Reading; Recording of previous events; Reproducibility; Research; Running; Sampling; Screening for Ovarian Cancer; Screening procedure; Sensitivity and Specificity; Serum; Solid; Speed; Staging; Structure; Surface; System; Time; Training; Translations; Tumor Antigens; Tumor Markers; Validation; base; cancer type; clinically relevant; combinatorial; design; falls; indexing; instrument; malignant breast neoplasm; particle; protein aminoacid sequence; serological marker; tumor

DESCRIPTION (provided by applicant): The development of circulating auto-antibodies to tumor-associated antigens (TAAs) has been observed at early cancer stages. TAA auto-antibodies are attractive as diagnostic markers because they are stable and persistent in cancerous conditions yet minimally present in normal individuals and most noncancerous conditions. However, the identification of auto-antibodies specific for a particular type of cancer is complicated due to the relatively low-abundance of an individual antibody within the complexity of the human proteome. The purpose of the present project is to develop an approach to detect auto-antibodies to tumor-associated antigens using an encoded combinatorial peptide library synthesized on PharmaSeq's light-activated radio-frequency p-Chips. Preparing the combinatorial library on the p-Chip platform serves two purposes: 1) to enrich for low-abundance proteins based on established principles of solid-phase affinity adsorption and 2) to rapidly identify the affinity ligand on each chip based on the encoded ID. The main project goal is to synthesize an RFID-encoded peptide library consisting of several thousands of random tetramers using the "split-and-mix" method. We will use a high speed fluidics-based analytical instrument previously developed by PharmaSeq to identify p-Chips carrying specific peptides with affinity to human auto- antibodies. In addition we will quantitatively characterize differences in immunoglobulin profiles between early stage ovarian and breast cancer patient and normal control samples. The methods developed will enable, for the first time, a true encoded one- particle-one-compound high throughput library synthesis and screening method that is capable of direct translation as a clinical diagnostic platform. PUBLIC HEALTH RELEVANCE: Auto-antibodies generated against tumor-associated antigens show great potential for the accurate, early diagnosis of cancer but identifying antibodies specific to a particular type of cancer is challenging, requiring a sensitive, high-throughput system that can discriminate key biomarkers against a high protein load. The implementation of an RFID- encoded combinatorial peptide library with PharmaSeq's light-activated p-Chip system will enable the simultaneous enrichment and identification of clinically-relevant biomarkers on a unified, high-throughput platform.

描述（由申请人提供）：在早期癌症阶段已经观察到循环自身抗体向肿瘤相关抗原（TAA）的发展。 TAA自身抗体作为诊断标记具有吸引力，因为它们在癌状态下稳定且持续存在，但在正常个体和大多数非癌状况下都最少地存在。但是，鉴定特定于特定类型癌症的自身抗体 由于人类蛋白质组复杂性内的个体抗体相对较低，因此很复杂。本项目的目的是开发一种方法，使用在Pharmaseq的光激活射频P-CHIP上合成的编码组合肽库来检测与肿瘤相关抗原的自身抗体。在P-Chip平台上准备组合文库有两个目的：1）基于固定相位亲和力吸附的既定原理，以富含低丰度蛋白质的浓缩蛋白，2）根据编码ID快速识别每个芯片上的亲和力配体。主要的项目目标是合成RFID编码的肽库，该肽库由数千种随机四聚体使用“拆分和混合”方法组成。我们将使用先前由Pharmaseq开发的基于高速流体的分析仪器来识别具有与人类自动抗体亲和力的特定肽的P芯片。此外，我们将定量地表征早期卵巢癌和乳腺癌患者和正常对照样品之间免疫球蛋白特征的差异。开发的方法将首次启用真正的编码单粒子单粒子高吞吐量库合成和筛选方法，该方法能够直接翻译为临床诊断平台。 公共卫生相关性：针对肿瘤相关抗原产生的自身抗体表现出很大的潜力，可以对癌症进行准确的早期诊断，但是识别特定类型癌症的抗体具有挑战性，需要敏感，高通量的系统，可以将关键生物标志物区别于高蛋白质负荷。使用Pharmaseq的光激活P-CHIP系统的RFID编码组合肽库的实施将使在统一的高通量平台上同时富集和鉴定临床上相关的生物标志物。