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

在这里，我们提出了一种突破性的技术，将药物靶受体呈现在纳米颗粒上，用于核酸适体的生物特异性纯化，即特异性肽药物，可以通过液相色谱电喷雾电离和串联质谱（LC-ESI-MS/MS）的分析能力以及64位SQL Server-R计算进行鉴定和定量。像IgG这样的免疫球蛋白抗体是一类特异性结合血液配体或细胞表面受体（如与癌症等疾病相关的受体）的生物药物。IgG等循环蛋白配体与细胞表面受体靶点结合，可作为高度特异性的生物药物治疗疾病。迫切需要将配体（如蛋白质、抗体、核酸或其受体）呈现在致密的亲水二氧化硅纳米颗粒上，以分离配体-受体复合物，并通过强大的串联质谱（LC-ESI-MS/MS）进行分析，从而发现新的药物和新的药物靶点。此外，还需要滴定循环中特定分子的浓度，并使用功能强大的LC-ESI-MS/MS或超灵敏的酶联质谱分析（ELiMSA）来量化它们与活细胞或固定细胞表面的结合。IgG抗体的特异性来自于可变结构域氨基酸的近乎随机重排。因此，使用随机肽适体或来自胎儿血清的天然肽作为原型生物药物来生产针对蛋白质药物靶点的生物药物可能会有飞跃。可溶配体，受体的可溶部分，或受体相关蛋白，可以作为药物靶标来筛选可能直接作为原型药物的肽适体的结合。相反，我们可以使用特定的配体，如二氧化硅纳米颗粒上的IgG来捕获可能成为治疗药物结合靶点的受体蛋白复合物。携带IgG或oxLDL配体的微颗粒已被用于捕获Fc和oxLDL受体，并鉴定和量化经药物和沉默RNA证实的新的信号因子。因此，我们认为，通过LC-ESI-MS/MS对二氧化硅纳米珠进行生物特异性纯化，可以分离出新的药物靶点，筛选和揭示新的高度特异性的生物多肽药物，并且可以使用ELiMSA快速、灵敏地滴定和定量其与细胞的结合，这将彻底改变我们对疾病的认识和治疗。

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.