Methods to Identify High-Affinity Antibodies that Target Tumor-Associated Glycans

鉴定针对肿瘤相关聚糖的高亲和力抗体的方法

基本信息

批准号：
8504989
负责人：
Jonathan R. Lai
金额：
$ 19.54万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8504989
关键词：
Affinity Amino Acids Animal Sources Animals Antibodies Antibody Affinity Antibody Diversity Antibody Formation Antigens Architecture Bacteriophages Benchmarking Binding Binding Sites Bioinformatics Biological Biological Process Cancer Diagnostics Case Study Cell surface Chemicals Cloning Codon Nucleotides Complementarity Determining Regions DNA Sequence Rearrangement Development Diagnosis Diagnostic Diagnostic Reagent Disease Dissociation Elements Evaluation Event Glycoproteins Goals HIV Envelope Protein gp120 HIV-1 Human Hybridomas Immunization Immunoglobulin G Individual Libraries Malignant Neoplasms Methodology Methods Monoclonal Antibodies Mus Nucleic Acids Oligonucleotides Oligosaccharides Peptides Phage Display Plant Roots Play Polysaccharides Positioning Attribute Production Property Proteins Randomized Reagent Reporting Research Role Source Specificity Surface Techniques Technology Therapeutic Ursidae Family Variant anticancer research base cancer therapy chemotherapy design gene function humanized antibody immunogenic innovation innovative technologies insight novel protein function scaffold synthetic construct targeted delivery tool tumor tumorigenesis vector

项目摘要

DESCRIPTION (provided by applicant): Monoclonal antibodies are essential reagents for deciphering gene or protein function and have been a fruitful source of therapeutic and diagnostic agents. However, the use of anticarbohydrate antibodies to target glycans for these purposes has been less successful. Glycans contain less hydrophobic functionality than do proteins or nucleic acids, thus individual glycan-antibody interactions are relatively weak. Information encoded by glycans often involves subtle variations of branched oligosaccharides that cannot be detected with conventional antibodies. It has therefore been difficult to obtain reagents that enable a complete understanding of biological glycan function. Changes in cell surface glycan composition are associated with cancer (and other disease states); therefore, general methods to identify specific and high- affinity glycan antibodies would greatly facilitate cancer research and provide new avenues for cancer therapies and diagnostics. We propose to develop methods to identify such reagents using novel antibody phage display technologies. It has recently become possible to select specific and high affinity antibodies for virtually any protein antigen from phage libraries that bear tailored diversity elements encoded by synthetic DNA ('synthetic antibodies'). This innovative technology platform obviates the requirement for animal immunization, thereby circumventing many limitations of traditional hybridoma methods. We will use the unique architectural scaffold of 2G12, an antibody that targets oligomannoses on the HIV-1 glycoprotein gp120, as the template for our design. The two heavy chain variable domains of 2G12 IgG exchange positions to create an extended recognition surface containing four oligomannose binding sites per IgG molecule. We have developed a phage vector that allows the functional display of the 2G12 scaffold. We will prepare synthetic 2G12 antibody libraries to determine minimal physicochemical requirements for high affinity glycan recognition in this scaffold. Next, we will use the information gained from these studies to identify novel 2G12 variants with altered specificity profiles for tumor-associated glycan targets. This innovative strategy will result in novel cancer antibodies that accelerate cancer research and pave the way for development of new cancer therapies and diagnostics.

描述（由申请人提供）：单克隆抗体是破译基因或蛋白质功能的重要试剂，并且是治疗和诊断试剂的丰富来源。然而，使用抗碳水化合物抗体来靶向聚糖以达到这些目的却不太成功。聚糖比蛋白质或核酸含有更少的疏水功能，因此单个聚糖-抗体相互作用相对较弱。聚糖编码的信息通常涉及支链寡糖的细微变化，而传统抗体无法检测到这些变化。因此，很难获得能够完全了解生物聚糖功能的试剂。细胞表面聚糖组成的变化与癌症（和其他疾病状态）有关；因此，鉴定特异性和高亲和力聚糖抗体的通用方法将极大地促进癌症研究，并为癌症治疗和诊断提供新途径。我们建议开发使用新型抗体噬菌体展示技术来识别此类试剂的方法。最近，从噬菌体文库中选择几乎任何蛋白质抗原的特异性和高亲和力抗体已成为可能，这些噬菌体文库带有由合成 DNA 编码的定制多样性元素（“合成抗体”）。这一创新技术平台无需动物免疫，从而规避了传统杂交瘤方法的许多限制。我们将使用 2G12 的独特结构支架（一种针对 HIV-1 糖蛋白 gp120 上的寡甘露糖的抗体）作为我们设计的模板。 2G12 IgG 的两个重链可变结构域交换位置，以创建每个 IgG 分子包含四个寡甘露糖结合位点的扩展识别表面。我们开发了一种噬菌体载体，可以实现 2G12 支架的功能展示。我们将制备合成 2G12 抗体库，以确定该支架中高亲和力聚糖识别的最低理化要求。接下来，我们将利用从这些研究中获得的信息来识别新的 2G12 变体，这些变体对肿瘤相关聚糖靶点具有改变的特异性。这一创新策略将产生新型癌症抗体，加速癌症研究，并为新癌症疗法和诊断的开发铺平道路。