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：
8294534
负责人：
Jonathan R. Lai
金额：
$ 20.79万
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/8294534
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必不可少的试剂，并且已成为治疗和诊断剂的富有成效的来源。然而，出于这些目的的抗碳水化合物抗体将抗体抗体靶向聚糖并不成功。与蛋白质或核酸相比，聚糖的疏水功能较少，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无法用常规抗体检测到的分支寡糖的细微变化。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，鉴定特定和高亲和力聚糖抗体的一般方法将极大地促进癌症研究，并为癌症疗法和诊断提供新的途径。我们建议使用新型抗体噬菌体显示技术开发方法来识别此类试剂。最近，可以从噬菌体文库中选择任何蛋白质抗原的特定和高亲和力抗体，这些蛋白质抗原具有由合成DNA（“合成抗体”）编码的量身定制的多样性元素。这个创新的技术平台避免了动物免疫的需求，从而规避了传统杂交方法的许多局限性。我们将使用2G12的独特建筑支架，该抗体靶向HIV-1糖蛋白GP120上的寡聚剂，作为我们设计的模板。 2G12 IgG交换位置的两个重链可变域，以创建一个扩展识别表面，该识别表面包含每个IgG分子的四个寡素糖结合位点。我们已经开发了一个噬菌体向量，该向量允许2G12支架的功能显示。我们将准备合成2G12抗体文库，以确定该支架中高亲和力识别的最小物理化学需求。接下来，我们将使用从这些研究中获得的信息来识别与肿瘤相关的聚糖靶标具有变化特异性曲线的新型2G12变体。这种创新的策略将导致新型的癌症抗体加速癌症研究，并为开发新的癌症疗法和诊断症铺平了道路。