Biophysical and biochemical techniques for the analysis and targeting of ligand-receptor supramolecular complexes

用于分析和靶向配体-受体超分子复合物的生物物理和生化技术

• 批准号: RGPIN-2019-05738
• 负责人: Marshall, John
• 金额: $ 1.75万
• 依托单位: Ryerson University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=679570
• 关键词: Biophysical; biochemical; techniques; analysis; targeting

Here we propose to create a breakthrough technology to present drug target receptors on nano particles for the bio-specific purification of aptamers, i.e. specific peptides drugs, that may be identified and quantified by the analytical power of liquid chromatography electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS), together with 64 bit SQL Server-R computation. Immunoglobulin antibodies like IgG are a class of biological drugs that specifically bind blood ligands, or cell surface receptors, such as the receptors associated with diseases including cancer. Circulating protein ligands such as IgG bind to cell surface receptor targets and may act as highly specific biological drugs in the treatment of disease. There is an urgent need to present ligands such as proteins, antibodies, nucleic acids, or their receptors, on dense, hydrophilic silica nano particles to isolate ligand receptor complexes for analysis by powerful tandem mass spectrometry (LC-ESI-MS/MS) to discover new drugs and new drug targets. Furthermore, there is a need to titre the concentration of specific molecules in circulation and to quantify their binding to the surface of live or fixed cells using powerful LC-ESI-MS/MS or ultra-sensitive enzyme linked mass spectrometric assays (ELiMSA). The specificity of IgG antibodies results from the near random rearrangement of amino acids in the variable domain. Thus it may be possible to leap forward in the production of biological drugs against a protein drug target using random peptide aptamers, or natural peptides from fetal serum, as prototypic biological drugs. A soluble ligand, the soluble portion of the receptor, or receptor associated proteins, may be presented as drug targets to screen the binding of peptide aptamers that might directly serve as prototypic drugs. Conversely, we may use specific ligands such as IgG presented on silica nano particles to capture the receptor protein complex that may be the binding target of therapeutic drugs. Micro particles carrying IgG or oxLDL ligands have been used to capture the Fc and oxLDL receptors and identify and quantify new signaling factors as confirmed by drugs and silencing RNA. Thus we propose that bio-specific purification on silica nano beads interrogated by powerful LC-ESI-MS/MS may isolate new drug targets, screen and reveal new and highly specific biological peptide drugs, and that ELiMSA may be used to rapidly and sensitively titre and quantify the binding to cells, that together will revolutionize our understanding and treatment of disease.

在这里，我们提出了一项突破性的技术，将药物靶标受体呈现在纳米粒子上，用于适体的生物特异性纯化，即特定的多肽药物，可以通过LC-ESI-MS/MS的分析能力和位SQL Server-R的计算来鉴定和定量。免疫球蛋白抗体，如免疫球蛋白抗体，是一类特定结合血液配体或细胞表面受体的生物药物，如与癌症等疾病相关的受体。循环蛋白配体如免疫球蛋白G与细胞表面受体靶点结合，在疾病治疗中可作为高度特异的生物药物。迫切需要将蛋白质、抗体、核酸或它们的受体等配体呈现在致密、亲水性的二氧化硅纳米颗粒上，以分离配体受体复合体，用于强大的串联质谱仪(LC-ESI-MS/MS)分析，以发现新药和新的药物靶点。此外，还需要使用强大的LC-ESI-MS/MS或超灵敏的酶联质谱(ELiMSA)来滴定循环中特定分子的浓度，并量化它们与活细胞或固定细胞表面的结合。免疫球蛋白抗体的特异性源于可变区氨基酸的近乎随机重排。因此，使用随机多肽适配子或来自胎儿血清的天然多肽作为原型生物药物，针对蛋白质药物靶标生产生物药物是可能的。一个可溶的配体，即受体的可溶部分，或受体相关蛋白，可以作为药物靶点来筛选可能直接作为原型药物的多肽适配子的结合。相反，我们可以使用特定的配体，如二氧化硅纳米颗粒上的免疫球蛋白来捕获可能是治疗药物结合靶点的受体蛋白复合体。携带免疫球蛋白或氧化低密度脂蛋白配体的微粒已被用于捕获Fc和ox低密度脂蛋白受体，并识别和量化新的信号因子，如药物和沉默RNA所证实的。因此，我们认为，高效LC-ESI-MS/MS对二氧化硅纳米球的生物特异性纯化可以分离新的药物靶点，筛选和揭示新的高度特异的生物肽药物，ELiMSA可以用于快速和灵敏地滴定和定量与细胞的结合，这些都将彻底改变我们对疾病的理解和治疗。