Design, expression, and properties of rHbs for HBOCs

HBOC 的 rHb 的设计、表达和特性

• 批准号: 6654243
• 负责人: CHIEN HO
• 金额: $ 38.74万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-05 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6654243
• 关键词: Bohr effect; Escherichia coli; biomaterial development /preparation; biotechnology; blood /plasma substitute; dextrans; fluidity; hemoglobin; nitric oxide; nitroso compounds; nuclear magnetic resonance spectroscopy; oxygen transport; peptide chemical synthesis; polyethylene glycols; protein engineering; protein structure function; proteomics; recombinant proteins

DESCRIPTION (provided by applicant): The goal of this project is to design and express novel human mutant recombinant hemoglobins (rHbs), which, after reacting with polyethylene glycol (PEG) or other cross-linking reagents, possess appropriate structural and functional properties to serve as hemoglobin-based oxygen carries (HBOCs). The vasoactivity associated with the use of many of the current HBOCs has challenged previous beliefs as to the optimal O2 affinity, cooperatively, Bohr effect, size, viscosity, and other properties needed for HBOCs. rHbs that we have previously prepared and new ones that we will design and express will be used to test both the new paradigms for the design of blood substitutes and the various hypotheses regarding the origin of vasoactivity. Using our Hb expression system in Escherichia coli, we are in a position to design and express any rHbs needed. Our laboratory has extensive experience and is also fully equipped to carry out structural studies using nuclear magnetic resonance (NMR) and functional studies using biochemical-biophysical techniques (e.g., equilibrium oxygen binding, kinetics of ligand binding, etc)on Hbs. A unique feature of this Program Project is the interactions among its component parts. The key investigators of this Program Project have worked together and have published papers together during the past ten years. The scope of this Program Project requires a multidisciplinary approach to the design of novel HBOCs. The structural and biochemical-biophysical results obtained from this project (Project 2) will provide new insights for Projects 1, 3, 4 and 5 as well as the Protein Biochemistry Core. Equally, the results from other components of the Program Project will guide Project 2 in designing the rHbs most effective as HBOCs. We believe that the results obtained from this Program Project will provide new knowledge for the development of safe, reliable HBOCs as blood substitutes.

描述（由申请人提供）： 该项目的目的是设计和表达新型的人类突变体重组血红蛋白 （RHB），它与聚乙烯乙二醇（PEG）或其他交联试剂反应后 具有适当的结构和功能特性，可作为基于血红蛋白的氧气 携带（HBOC）。与使用许多当前HBOC有关的血管活性 挑战了先前关于最佳O2亲和力的信念，合作，bohr效应，大小， HBOC所需的粘度和其他特性。我们以前准备的RHB， 我们将设计和Express的新范式将用于测试 血液替代品的设计和有关血管活性起源的各种假设。 在大肠杆菌中使用我们的HB表达系统，我们可以设计和表达 任何RHB都需要。我们的实验室有丰富的经验，还可以携带 使用核磁共振（NMR）和功能研究进行结构研究 生化生物物理技术（例如，平衡氧结合，配体的动力学 绑定等）在HBS上。该程序项目的一个独特功能是其之间的互动 组件零件。该计划项目的关键调查人员共同努力， 在过去的十年中，已经一起发表了论文。该计划项目的范围 需要一种多学科的方法来设计新型HBOC。结构和 从该项目获得的生化生物物理结果（项目2）将为项目1、3、4和5以及蛋白质生物化学核心提供新的见解。同样，来自其他的结果 该计划项目的组件将指导项目2设计最有效的RHB HBOC。我们认为，从该计划项目获得的结果将提供新的 了解安全，可靠的HBOC作为血液替代品的知识。