Isolation of Monoclonal Antibodies against Membrane Proteins

描述（由申请人提供）：由于其在细胞信号传导和人类疾病中的重要性，整合膜蛋白如G蛋白偶联受体（GPCR）和离子通道占现有药物靶标的40%以上。识别膜蛋白上的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，因为它们通常与可用于其检测或抑制的受体的关键结构结合。然而，构象依赖性的，抑制性单克隆抗体对膜蛋白的发展是特别困难的，因为，不像可溶性蛋白，大多数膜蛋白依赖于脂质环境，以保持其天然的三级和四级（寡聚）结构。膜蛋白通常难以纯化，通常在细胞表面上以低浓度表达，并且通常很少由线性肽表示。需要新的方法来开发用于治疗、诊断和研究应用的此类单克隆抗体。在这里，我们建议使用一种新的技术，Lipoparticle，捕获和浓缩结构完整的膜蛋白的格式服从免疫。使用Lipoparticles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使用的基础上。 公共卫生相关性：该提案将导致针对重要膜蛋白靶点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。

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.