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

• 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
• 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

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(申请人提供)：单抗是破译基因或蛋白质功能的基本试剂，是治疗和诊断试剂的丰富来源。然而，为了这些目的而使用抗碳水化合物抗体来靶向糖链的研究就不那么成功了。与蛋白质或核酸相比，葡聚糖含有较少的疏水功能，因此单个糖链与抗体的相互作用相对较弱。用多聚糖编码的信息通常涉及用常规抗体无法检测到的分枝低聚糖的细微变化。因此，很难获得能够完全了解生物多糖功能的试剂。细胞表面糖链组成的变化与癌症(和其他疾病状态)有关；因此，鉴定特异性和高亲和力的糖链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的途径。我们建议开发使用新型抗体噬菌体展示技术来鉴定这类试剂的方法。最近已经有可能从带有由合成DNA(合成抗体)编码的定制多样性元件的噬菌体库中选择针对几乎任何蛋白质抗原的特异性和高亲和力抗体。这一创新的技术平台避免了对动物免疫的要求，从而绕过了传统杂交瘤方法的许多局限性。我们将使用2G12的独特结构支架作为我们设计的模板，2G12是一种针对HIV-1糖蛋白gp120上的寡聚注释的抗体。2G12免疫球蛋白的两个重链可变区交换位置，形成一个延伸的识别表面，每个免疫球蛋白分子包含四个寡核苷酸结合部位。我们开发了一种噬菌体载体，可以展示2G12支架的功能。我们将准备合成的2G12抗体库，以确定在这种支架上识别高亲和力多糖所需的最低物理化学要求。接下来，我们将使用从这些研究中获得的信息来识别具有肿瘤相关糖链靶标特异性改变的新型2G12变异体。这一创新战略将产生新的癌症抗体，加速癌症研究，并为新的癌症治疗和诊断方法的开发铺平道路。

项目成果

A strategy for phage display selection of functional domain-exchanged immunoglobulin scaffolds with high affinity for glycan targets.

噬菌体展示选择对聚糖靶标具有高亲和力的功能域交换免疫球蛋白支架的策略。

• DOI: 10.1016/j.jim.2011.12.008
• 发表时间: 2012
• 期刊: Journal of immunological methods
• 影响因子: 2.2
• 作者: Stewart,Alex;Liu,Yanyun;Lai,JonathanR
• 通讯作者: Lai,JonathanR