Novel Single Domain Antibodies with Multivalency and Multispecificity

具有多价性和多特异性的新型单域抗体

基本信息

批准号：
8399006
负责人：
Rihe Liu
金额：
$ 28.74万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
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：
2011-12-09 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8399006
关键词：
Address Affinity Amino Acid Sequence Amino Acids Animal Model Animals Antibodies Apoptosis Avidity Bacteria Binding Biological Products Breast Calmodulin Cell Proliferation Chemicals Collaborations Colorectal Complex Cytoplasm Development Diagnosis Diagnostic Disadvantaged Drug Delivery Systems ERBB2 gene ERBB3 gene Engineering Epidermal Growth Factor Receptor Epitopes Evolution Extracellular Domain Family Family member Gene Delivery Generations Genes Head and Neck Cancer Human Image In Vitro Interdisciplinary Study Investigation Lead Length Libraries Ligands Lung Malignant Neoplasms Malignant neoplasm of pancreas Messenger RNA Methods Mono-S Nanotechnology Neoplasm Metastasis Pancreatic Ductal Adenocarcinoma Patients Penetration Peptide Sequence Determination Peptides Plant Leaves Play Post-Translational Protein Processing Proteins Receptor Protein-Tyrosine Kinases Regulation Research Role SHFM1 gene Signal Transduction Site Small Interfering RNA Specificity Structure Surgical Oncologist System Technology Tertiary Protein Structure Therapeutic Agents Time Tissues Xenograft procedure base calcium phosphate cancer cell cancer diagnosis cancer therapy cancer type cell motility cost cost effective directed evolution humanized monoclonal antibodies immunogenicity in vivo interest malignant stomach neoplasm member mouse model nanoparticle neoplastic cell next generation novel receptor scaffold self assembly success targeted delivery tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): ErbB/HER receptors play essential roles in propagating signals that regulate cell proliferation, differentiation, motility, and apoptosis. Significantly, aberrant regulation of the ErbB/HER family of receptor tyrosine kinases and their ligands is a common occurrence in many human tumors. Indeed, EGFR, HER2, and HER3 have been diagnostically and therapeutically validated as targets in a wide variety of human malignancies such as lung, colorectal, breast, head and neck, gastric and pancreatic cancers. Therefore, novel protein biopharmaceuticals that comprehensively target ErbB/HER family members could have significant applications in targeted diagnosis and therapy of different types of cancer. This project addresses a critical need for developing cost-effective targeting ligands that recognize and bind HER family members with desired avidity and multispecificity. The project is directed at using a powerful in vitro protein selection method called mRNA display to evolve single domain antibodies (SDAs) with human origin that tightly and specifically bind to the extracellular domain of EFRG, HER2 or HER3, respectively, from a human SDA domain library with an unusually high diversity. The resulting monomeric SDAs will be used as the basis for the generation of heterodimeric SDAs that bind to a HER member of interest at two nonoverlapping epitopes with synergistic avidity effect. In addition, we will develop an efficient and universal self-assembly system that allows for reversible and controllable loading of HER-binding SDAs to nanoparticles to acquire desired multispecificity. The resulting SDAs would have a number of advantages, including high target-binding affinity, ease to achieve desired avidity, multivalency and multispecificity, minimal immunogenicity due to human origin, small size for better tissue penetration, in addition to significantly reduced manufacturing costs due to high expression levels in bacteria. The simplicity and stability of these SDAs also greatly facilitate their conjugation with various nanoparticles for translational applications. The resulting SDAs will be examined in vivo in pancreatic ductal adenocarcinoma (PDAC) xenograft animal models by targeted delivery of nanoparticles containing siRNAs against several metastasis signature genes that are important for PDAC tumorigenesis and metastasis. The success of the project will lead to the development of cost-effective, targeted biopharmaceuticals for cancer diagnosis and therapy.

描述（由申请人提供）：ERBB/她的受体在传播调节细胞增殖，分化，运动性和凋亡的信号中起着重要作用。值得注意的是，在许多人类肿瘤中，ERBB/她的受体酪氨酸激酶家族及其配体的异常调节是很常见的。的确，EGFR，HER2和HER3在肺部恶性肿瘤（例如肺，结肠直肠，乳房，头颈，胃和胰腺癌）等各种人类恶性肿瘤中已被诊断和治疗验证。因此，全面针对ERBB/她的家人的新型蛋白质生物药物可以在针对性诊断和治疗不同类型的癌症的靶向诊断和治疗中有重要应用。该项目解决了建立具有成本效益的靶向配体的关键需求，这些配体以所需的亲戚和多特殊性来识别和束缚她的家人。该项目旨在使用称为mRNA显示的功能强大的体外蛋白质选择方法，以分别与人类SDA域库的EFRG，HER2或HER3的细胞外域分别与EFRG，HER2或HER3的细胞外域紧密结合，并与具有异常高的多样性的人类SDA域库中分别与EFRG，HER2或HER3的细胞外域进行紧密结合。所得的单体SDA将被用作生成异二聚体SDA的基础，这些杂种SDA与她的感兴趣成员在两个非重叠的表位具有协同伴随的效果。此外，我们还将开发一个有效且通用的自组装系统，该系统允许将HER结合SDA的可逆且可控制的加载到纳米颗粒中获得所需的多特异性。由此产生的SDA将具有许多优势，包括高目标结合亲和力，易于实现所需的亲和力，多类和多特异性，由于人类起源而引起的最低免疫原性，除了由于细菌的高表达水平而显着降低了生产成本，因此较小的组织尺寸较小。这些SDA的简单性和稳定性也极大地促进了它们与各种纳米颗粒的共轭，用于转化应用。将在胰腺导管腺癌（PDAC）异种移植动物模型中在体内检查所得的SDA，通过针对含有siRNA的纳米颗粒的靶向递送，这些纳米颗粒针对几个转移性签名基因，这些基因对PDAC肿瘤发生和转移很重要。该项目的成功将导致开发具有成本效益的靶向生物药物来进行癌症诊断和治疗。