Isolation of Monoclonal Antibodies against Membrane Proteins

膜蛋白单克隆抗体的分离

基本信息

批准号：
8215713
负责人：
JOSEPH Benjamin RUCKER
金额：
$ 33.83万
依托单位：
INTEGRAL MOLECULAR
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：
2011-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8215713
关键词：
Abbreviations Adjuvant Antibodies Antigens Binding Biomedical Research Cell Line Cell surface Cells Cellular Membrane DNA Detection Detergents Development Diagnostic Diagnostics Research Drug Delivery Systems Environment Epitopes G-Protein-Coupled Receptors Goals Hybridomas Immune Immune response Immunization Immunologic Adjuvants Integral Membrane Protein Ion Channel Licensing Lipid Bilayers Lipids Marketing Membrane Membrane Proteins Methods Molecular Conformation Monoclonal Antibodies Mus Peptides Preparation Production Property Proteins Protocols documentation Reagent Research Serum Signal Transduction Source Structure Testing Therapeutic Monoclonal Antibodies TimeLine Vaccines Viral Viral Vaccines Virus-like particle expression vector human PHEMX protein human disease improved new technology novel strategies particle product development public health relevance receptor receptor function success therapeutic development vaccine development

项目摘要

DESCRIPTION (provided by applicant): Because of their importance in cell signaling and human disease, integral membrane proteins such as G protein-coupled receptors (GPCRs) and ion channels comprise over 40% of existing drug targets. Monoclonal antibodies (MAbs) that recognize conformation-dependent epitopes on membrane proteins are usually the most valuable type of antibody because they often bind to critical structures of the receptor that can be exploited for its detection or inhibition. However, the development of conformation-dependent, inhibitory MAbs against membrane proteins is especially difficult because, unlike soluble proteins, most membrane proteins are dependent on a lipid environment to maintain their native tertiary and quaternary (oligomeric) structures. Membrane proteins are often difficult to purify, often express at low concentrations on the cell surface, and are usually poorly represented by linear peptides. New approaches are needed to develop such MAbs for therapeutic, diagnostic, and research applications. Here we propose to use a novel technology, the Lipoparticle, to capture and concentrate structurally intact membrane proteins in a format amenable to immunization. The concept of using Lipoparticles to develop antibodies against cellular membrane proteins builds on the historic use of viral particles as successful vaccines. PUBLIC HEALTH RELEVANCE: This proposal will result in monoclonal antibodies against important membrane protein targets for therapeutic development, diagnostics, and biomedical research. Lipoparticles optimized as immunogens will be developed as commercial products.

描述（由申请人提供）：由于 G 蛋白偶联受体 (GPCR) 和离子通道等整合膜蛋白在细胞信号传导和人类疾病中的重要性，它们构成了现有药物靶标的 40% 以上。识别膜蛋白上构象依赖性表位的单克隆抗体 (MAb) 通常是最有价值的抗体类型，因为它们通常与受体的关键结构结合，可用于检测或抑制受体。然而，针对膜蛋白的构象依赖性、抑制性单克隆抗体的开发尤其困难，因为与可溶性蛋白不同，大多数膜蛋白依赖于脂质环境来维持其天然的三级和四级（寡聚）结构。膜蛋白通常难以纯化，通常在细胞表面以低浓度表达，并且通常很难用线性肽表示。需要新的方法来开发此类单克隆抗体用于治疗、诊断和研究应用。在这里，我们建议使用一种新技术，即脂质颗粒，以适合免疫的形式捕获和浓缩结构完整的膜蛋白。使用脂质颗粒开发针对细胞膜蛋白的抗体的概念建立在病毒颗粒作为成功疫苗的历史用途之上。公共健康相关性：该提案将产生针对重要膜蛋白靶标的单克隆抗体，用于治疗开发、诊断和生物医学研究。作为免疫原优化的脂质颗粒将被开发为商业产品。