Jefferson Shared Biacore Instrumentation

描述(由申请人提供)：托马斯·杰斐逊大学坎摩尔癌症中心的核心用户群体提议购买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(亲和力传感器)作为分子相互作用装置的核心仪器来测量生物分子相互作用的动力学。虽然这种仪器已经使用户能够成功地执行一些项目，但新的和更具挑战性的实验将需要更复杂的仪器。 Biacore 3000提供的噪音比IAsys低约10倍，具有更高的灵敏度，并允许检测低分子配体的结合，而不仅仅是大分子。Biacore 3000还提供更精确的温度稳定性(q0.IxC比q0.30℃)和2倍宽的温度范围，这对于精确的生物分子相互作用的热力学分析是必不可少的。Biacore 3000能够测量最宽范围的折射率，并拥有卓越的Global数据分析软件。此外，我们目前在分子相互作用设施中拥有的单一仪器不能适应不断增长的用户数量。 有了像Biacore 3000这样的现代生物传感器，核心用户将能够全面地解决以下问题：1.根据抗原特异性受体与同源抗原反应的热力学，什么分子机制决定了抗原特异性受体的特异性？它们机制的不同将与B细胞和T细胞受体识别的抗原宇宙的不同有关；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癌变；3.蛋白质-蛋白质和蛋白质-DNA相互作用如何指导定点基因转位？了解这一过程的准确性是应用定点转座进行基因治疗的先决条件；4.5-螺旋和C-肽抑制剂如何与gp41相互作用以阻止HIV-1进入？我们必须了解它们的机制才能设计稳定的小分子疗法；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的影响？我们必须了解配体-受体的相互作用，以验证新的靶点，并确定新的治疗方法。

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.