Jefferson Shared Biacore Instrumentation

描述（由申请人提供）：托马斯杰斐逊大学Kimmel癌症中心的核心用户组提出购买BIAcore 3000表面等离子体共振生物传感器。我们一直使用IAsys共振镜生物传感器（亲和传感器）作为分子相互作用设施的核心仪器来测量生物分子相互作用的动力学。虽然该仪器已使用户成功地执行了一些项目，但新的和更具挑战性的实验将需要更复杂的仪器。 BIAcore 3000的噪音比IAsys低约10倍，灵敏度更高，不仅可以检测大分子，还可以检测低分子量配体的结合。BIAcore 3000还提供更精确的温度稳定性（q0.1xC与q0.30C最好）和2倍宽的温度范围，这对于生物分子相互作用的精确热力学分析至关重要。BIAcore 3000能够测量最宽范围的折射率，并具有用于数据分析的上级Global软件。此外，我们目前在分子相互作用设施中拥有的单一仪器无法容纳越来越多的用户。 通过BIAcore 3000等现代生物传感器，核心用户将能够全面解决以下问题：1.什么样的分子机制决定了抗原特异性受体的特异性，正如它们与同源抗原反应的热力学所揭示的那样？其机制的差异将与B细胞和T细胞受体识别的抗原的差异有关; 2.蛋白质-蛋白质和蛋白质-DNA相互作用如何指导DNA修复？我们必须确定DNA修复基因的突变如何允许致癌; 3.蛋白质-蛋白质和蛋白质-DNA相互作用如何指导位点特异性基因转座？了解这一过程的准确性是将位点特异性转座应用于基因治疗的先决条件; 4. 5-Heptane和C肽抑制剂如何与gp 41相互作用以阻断HIV-1进入？我们必须了解它们的机制，以便设计稳定的小分子治疗剂; 5。环肽配体与其受体之间的相互作用是否有助于解释这些配体对细胞生理学的影响？我们必须了解配体-受体相互作用，以验证新的靶点和表征新的治疗方法。

DESCRIPTION (provided by applicant): The core group of users at the Kimmel Cancer Center of Thomas Jefferson University proposes to acquire a BIAcore 3000 surface plasmon resonance biosensor. We have been using an IAsys Resonant Mirror Biosenser (Affinity Sensor) as the core instrument of the Molecular Interactions Facility to measure the kinetics of biomolecular interactions. While this instrument has allowed the users to execute successfully a number of projects, new and more challenging experiments will require a more sophisticated instrument. The BIAcore 3000 provides approximately 10-fold lower noise than the IAsys, yielding much greater sensitivity and permitting detection of the binding of low molecular weight ligands, not just macromolecules. The BIAcore 3000 also provides more precise temperature stabilization (q0.IxC vs. q0.30C at best) and 2-times wider temperature range, which is essential for accurate thermodynamic analysis of biomolecular interactions. The BIAcore 3000 is capable of measuring the widest range of refractive indexes and has superior Global software for data analysis. In addition, the single instrument we currently have in the Molecular Interaction Facility cannot accommodate the growing number of users. With a modem biosensor such as the BIAcore 3000, the core users will be able to address comprehensively the following questions: 1. What molecular mechanisms dictate the specificity of antigen-specific receptors, as revealed by the thermodynamics of their reactions with cognate antigens? Differences in their mechanisms will be related to differences in the universe of antigens recognized by B-cell and T-cell receptors; 2. How do protein-protein and protein-DNA interactions guide DNA repair? We must determine how mutations in DNA repair genes permit carcinogenesis; 3. How do protein-protein and protein-DNA interactions guide site-specific gene transposition? Understanding the accuracy of this process is a prerequisite to applying site-specific transposition for gene therapy; 4. How do 5-Helix and C-peptide inhibitors interact with gp41 to block HIV-1 entry? We must understand their mechanisms in order to design stable small molecule-therapeutics; 5. Can the interactions between cyclic peptide ligands and their receptors help to explain the effect of those ligands on cell physiology? We must understand ligand-receptor interactions to validate new targets and characterize new therapeutics.