INSTRUMENT TO CHARACTERIZE CELL-FREE 0XYGEN CARRIERS

一种用于表征无细胞血红蛋白和/或无细胞血红蛋白和红细胞混合物的氧气转运的新仪器已经发明，并已申请专利。该仪器使用小样本测量扩散氧的传输，适用于无细胞血红蛋白溶液的开发和对接受该溶液的患者的监测。这项提议的目标是优化该仪器的设计，以使用人工毛细管系统测量体外无细胞血红蛋白溶液的氧气转运。在特定目标1下，我们将建立一个人工毛细管系统来评估体外无细胞血红蛋白溶液对O2的转运。我们将优化我们现有人工毛细管仪的面板设计。氧气转运的测量将使用无细胞血红蛋白溶液，包括聚乙二醇化血红蛋白和交联四聚体血红蛋白的模型溶液，以及红细胞悬浮液。在具体目标2下，我们将使毛细管仪器自动化，并进一步开发数据解释软件。数据分析模型使用了基于克罗圆柱体模型的数学概念，用于将氧气从定义的几何形状的圆柱管(即人造毛细管)中传输出来。该程序需要1)优化运行时间效率和2)根据人工毛细管设备中的血红蛋白溶液和/或红细胞的实验数据进行测试。这一提议背后的假设是，在对候选产品进行准确评估之前，需要一种新的方法来评估无细胞氧气载体的氧气传输。这是由于在无细胞载体存在的情况下O2的促进扩散现象，这可能导致自动调节的血管收缩。建议的商业应用：在无细胞氧气载体存在的情况下，氧气转运(Hb，Het)的临床评估方法是不充分的。我们已经发明了一种新的仪器来表征无细胞血红蛋白的氧气运输，其中包括促进扩散的评估。该仪器在接受了以血红蛋白为基础的产品的患者的临床测量中具有巨大的潜力，并且在测试新的候选溶液方面也具有潜力。

A new instrument to characterize O2 transport by cell-free hemoglobin and/or mixtures of cell-free hemoglobin and red blood cells has been invented, and a patent has been applied for. This instrument measures diffusional O2 transport using a small sample, and is applicable both to the development of cell-free hemoglobin solutions and to monitoring patients who have received them. The goal of this proposal is to optimize the design of this instrument to measure O2 transport by cell-free hemoglobin solutions in vitro using an artificial capillary system. Under Specific Aim 1, we will build an artificial capillary system to assess O2 transport by cell- free hemoglobin solutions in vitro. We will optimize the breadboard design of our present artificial capillary instrument. Measurements of O2 transport will be made using cell-free hemoglobin solutions, including model solutions of PEG-hemoglobin and cross-linked tetrameric hemoglobin, and RBC suspensions. Under Specific Aim 2, we will automate the capillary apparatus and further develop software for data interpretation. The data- analysis model uses mathematical concepts based on the Krogh cylinder model for O2 transport out a cylindrical tube of defined geometry (i.e., the artificial capillary). This programs requires 1) optimization for run time efficiency and 2) testing against experimental data from hemoglobin solutions and/or red blood cells in the artificial capillary device. The hypothesis underlying this proposal is that a new method to assess O2 transport by cell-free oxygen carriers is required before candidate products can be evaluated accurately. This is due to the phenomenon of facilitated diffusion of O2 in the presence of cell-free carriers, which can lead to autoregulatory vasoconstriction. PROPOSED COMMERCIAL APPLICATIONS: Methods of clinical evaluation of O2 transport (Hb, Het) in the presence of cell-free oxygen carriers are inadequate. We have invented a new instrument to characterize O2 transport by cell-free hemoglobin that includes evaluation of facilitated diffusion. This instrument has great potential for clinical measures in patients who have received hemoglobin- based products, as well as potential in the testing of new candidate solution.