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）： 该项目的目标是设计和表达新型人类突变重组血红蛋白 (rHbs)，与聚乙二醇(PEG)或其他交联剂反应后， 具有适当的结构和功能特性，可作为血红蛋白氧 携带（HBOC）。与当前许多 HBOC 的使用相关的血管活性已经 挑战了以前关于最佳 O2 亲和力、合作、玻尔效应、尺寸、 粘度以及 HBOC 所需的其他特性。我们之前准备好的 rHb 和 我们将设计和表达的新范式将用于测试新范式 血液替代品的设计以及有关血管活性起源的各种假设。 利用我们在大肠杆菌中的 Hb 表达系统，我们能够设计和表达 任何需要的rHb。我们的实验室拥有丰富的经验，也配备齐全 使用核磁共振 (NMR) 进行结构研究，并使用 生化生物物理技术（例如，平衡氧结合、配体动力学 Hbs 上的结合等）。该计划项目的一个独特之处在于其之间的相互作用 组成部分。该计划项目的主要研究者共同努力， 在过去的十年里共同发表了论文。本计划项目范围 需要采用多学科方法来设计新颖的 HBOC。结构和 从该项目（项目 2）获得的生物化学-生物物理结果将为项目 1、3、4 和 5 以及蛋白质生物化学核心提供新的见解。同样，其他的结果 该计划项目的组成部分将指导项目 2 设计最有效的 rHb HBOC。我们相信，该计划项目取得的成果将提供新的 开发安全、可靠的 HBOC 作为血液替代品的知识。