High level expression and purification of therapeutic human and mouse neuroserpin

治疗性人和小鼠神经丝氨酸蛋白酶抑制剂的高水平表达和纯化

• 批准号: 7393483
• 负责人: Duane Ellington Day
• 金额: $ 11.68万
• 依托单位: MEDIGENIX, LLC
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-15 至 2010-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7393483
• 关键词: Adjuvant; Animal Model; Animals; Binding; Biological; Cause of Death; Cell Line; Cells; Complex; Data; Development; Endopeptidases; Engineering; Equipment; Evaluation; Funding; Future; Grant; Human; Human Activities; In Vitro; Insecta; Intellectual Property; Ischemic Stroke; Kringles; Laboratories; Methodology; Methods; Michigan; Modeling; Molecular; Mus; Peptide Hydrolases; Phase; Phase II Clinical Trials; Plasmin; Plasminogen; Positioning Attribute; Production; Protease Inhibitor; Proteins; Protocols documentation; Purpose; Reagent; Recombinants; Recovery; Research; Resources; Rights; Site; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Stroke; System; Testing; Therapeutic; Therapeutic Agents; Tobacco; Treatment Efficacy; Universities; Virus; attenuation; base; commercialization; day; design; disability; in vitro Assay; in vivo; innovation; neuron loss; neuroserpin; novel; novel therapeutics; pre-clinical; programs; receptor binding; success; technology development

DESCRIPTION (provided by applicant): This Phase I SBIR proposal will investigate the feasibility of using a unique fusion tag system for the high level expression and purification of native human and mouse neuroserpin. Our collaborators at the University of Michigan have shown in animal models that the neuroserpin protein appears to have profound neuroprotective effects and therefore has great therapeutic potential in the treatment of ischemic stroke. To date, the material has proven to be exceeding difficult to obtain in the quantities necessary to conduct more detailed animal studies and the lack of the reagent has created a bottleneck in the development of more detailed and elaborate studies. We propose to produce the neuroserpin protein reagents in a stable insect cell line on a large scale and to subsequently test them in an in vitro assay for functional biological activity. These proteinase inhibitors will form the basis of an eventual Phase II study where they will be used therapeutically in a mouse stroke model. This model will allow for the evaluation of their efficacy in the attenuation of neuronal cell death associated with induced stroke. If successful, the new fusion tag system will also have a broader scope than just this proposal. This new tag will be very be generalized and have the potential to be commercialized for the purification of other important academic and industrial protein based reagents. Neuroserpin protein represents a promising new therapeutic in the treatment of ischemic stroke, which is the second leading cause of death and disability throughout the world. Medigenix LLC proposes to develop a new method for the production of this protein on a large scale. This methodology may also prove useful in many other protein-based therapeutics in the future.

描述（由申请人提供）：本I期SBIR提案将研究使用独特融合标签系统高水平表达和纯化天然人和小鼠神经丝氨酸蛋白酶抑制剂的可行性。我们在密歇根大学的合作者在动物模型中表明，neuroserpin蛋白似乎具有深刻的神经保护作用，因此在治疗缺血性中风方面具有巨大的治疗潜力。迄今为止，已证明很难获得进行更详细的动物研究所需数量的材料，而且试剂的缺乏已成为开展更详细和详细研究的瓶颈。我们建议在稳定的昆虫细胞系中大规模生产neuroserpin蛋白试剂，并随后在体外试验中测试它们的功能生物活性。这些蛋白酶抑制剂将形成最终的II期研究的基础，其中它们将用于小鼠中风模型的治疗。该模型将允许评估它们在与诱导的中风相关的神经元细胞死亡的衰减中的功效。如果成功，新的融合标签系统也将有一个更广泛的范围不仅仅是这个建议。这种新的标签将是非常普遍的，并有可能被商业化用于其他重要的学术和工业蛋白质基试剂的纯化。神经丝氨酸蛋白酶抑制剂是治疗缺血性卒中的一种很有前途的新疗法，缺血性卒中是世界上第二大致死和致残原因。Medigenix LLC建议开发一种大规模生产这种蛋白质的新方法。这种方法在未来也可能被证明在许多其他基于蛋白质的治疗中是有用的。