Human-derived hemoglobin based oxygen carrier for utilization in organ and tissue preservation

该项目旨在开发一种人源性血红蛋白氧载体(HBOC)溶液(第一阶段) 用于需要氧气输送溶液的多种体外应用(第二阶段)：1) 用于器官和组织保存的机器灌流(MP)系统，2)用于发现 生物标志物，以及3)在人类微芯片中灌流以用于药物发现。这是第一个开发的HBOC 特别适用于体外应用，该新方法是一种平衡体外液体的赋形剂 轮班。此外，这一新产品将在精简、高效的商业环境中开发 该流程将允许无缝过渡到具有成本效益的合同制造组织(CMO)。 VirTech Bio的HBOC和制造工艺是在成功进行体外概念验证后构思的 研究1使用HBOC(血液纯净®)。不幸的是，这家公司(OPK Biotech)于2014年破产。 血液净化的制造工艺复杂而昂贵，2，3排除了任何CMO的使用。这个 主要问题涉及牛的污染、氢气毒性以及是否需要昂贵和 专门的设备。VirTech Bio为这项SBIR第一阶段提案开发的新流程解决了所有 这些担忧的同时创造了更具成本效益的商业生产这一组成部分。 人类血红蛋白分子的最初来源将是通过过程4获得的过期人类血液 已经得到监管机构的批准。过期的人类红血球将通过超滤被“清洗”， 渗透冲击裂解，三级切向流过滤系统净化，去除细胞 杂质。随后，将与戊二醛聚合，然后进行席夫碱稳定化。这个 具体目标包括：1)生物物理表征以证明与原始HBOC的相似性，2) 分子量和储存参数的稳定性测试以及3)与机器的兼容性测试 在相关条件下灌流(无器官)12小时。 该产品一旦开发和验证，将在第二阶段与机器灌流装置相结合，并 根据我们以往的经验，用于肝脏保存。用于器官的新型HBOC 保存对临床移植有重要的积极影响。有效器官氧合 体外将提高目前器官保存的标准，并增加移植的数量。 应该有额外的积极的下游影响，通过减少发病率和死亡率 移植等待名单6，改善术后结果，减少住院时间7。 好处应该对降低临床移植的总体成本的能力有直接影响。 好了！

This project aims to develop a human derived Hemoglobin-Based Oxygen Carrier (HBOC) solution (Phase I) to be utilized in numerous ex-vivo applications (Phase II) where an oxygen delivery solution is required: 1) machine perfusion (MP) systems for organ and tissue preservation, 2) MP systems for the discovery of biomarkers, and 3) perfusate in human microchips for drug discovery. This is the first HBOC to be developed specifically for ex-vivo applications with the novel approach being an excipient that balances for ex-vivo fluid shifts. Furthermore, this new product will be developed within a streamlined, highly efficient commercial process that will allow a seamless transition to cost-effective Contract Manufacturing Organizations (CMO). VirTech Bio's HBOC and manufacturing process were conceived following successful proof-of-concept ex-vivo studies 1 with an HBOC (Hemopure®). Unfortunately, the company (OPK Biotech) went bankrupt in 2014. Hemopure had a complex and expensive manufacturing process 2,3 that precluded the use of any CMO. The main issues were related to bovine contamination, hydrogen gas toxicity and the need for expensive and specialized equipment. VirTech Bio's's new process developed for this SBIR Phase I proposal addresses all these concerns while creating a more cost-effective commercial production of this component. The initial source of human hemoglobin molecules will be expired human blood obtained through a process 4 already approved by regulatory bodies. The outdated human red blood cells will be `washed' via ultrafiltration, lysed by osmotic shock and purified by a three-step tangential flow filtration system to remove cellular impurities. Subsequently, polymerization will be with glutaraldehyde, followed by Schiff-base stabilization. The Specific aims include: 1) biophysical characterization to demonstrate similarity with the original HBOC, 2) stability testing of the molecular weight and storage parameters and 3) compatibility testing with machine perfusion (without the organs) for 12 hours at relevant conditions. This product once developed and validated will be coupled with a machine perfusion device in Phase II and utilized for liver preservation according to our previous experience. The new HBOC for utilization in organ preservation should have a significant positive impact in clinical transplantation. Effective organ oxygenation ex-vivo will enhance the current standards of organ preservation and increase the number of transplants 5. It should have additional positive downstream effects by decreasing the morbidity and the mortality on the transplant waiting list 6, improving post-operative outcomes and decreasing the length-of-stay 7. All these benefits should have a direct impact in the ability to decrease the overall costs in clinical transplantation 8. !