Murine Reagents and Analytical Tools

描述（由申请人提供）：通过采用各种实验技术和体内模型系统，已对控制血液凝固和纤维蛋白溶解的机制进行了广泛研究。最近，基因靶向（敲除小鼠）和转基因技术已被用于创建小鼠品系，其表型代表人类血栓性疾病和疾病状态。与此同时，在新药开发过程中通常使用鼠凝血模型。在开发新的促凝血、抗凝和溶凝药物的过程中，需要进行基础研发研究以及安全性、有效性和药代动力学研究。因为正常和基因改变的小鼠都是可用的，所以鼠模型是该应用的理想模型。基于鼠模型被普遍使用并且存在有限数量的试剂可用于辅助研究人员的事实，开发用于该研究领域的商业试剂组是必要的。该项目旨在通过产生一系列可用于人类系统的试剂和分析工具来提高小鼠凝血模型的实用性。长期目标是生产一系列可销售的鼠源性试剂，包括：a）用于血浆重建实验和测定标准化的纯化的鼠凝血因子; B）用于Western印迹、免疫组织化学、免疫沉淀、流式细胞术和ELISA应用的鼠凝血因子的抗体;和c）用于凝血酶原、酶、辅因子、抑制剂、活化肽和酶/抑制剂复合物的ELISA测定。利用传统的实验室技术，研究人员计划纯化广泛的止血蛋白，产生抗体，这些抗原的多克隆和单克隆抗体，并确定和开发具有临床重要意义的免疫测定与这些试剂。

DESCRIPTION (provided by applicant): The mechanisms that govern blood coagulation and fibrinolysis have been studied extensively by employing a variety of experimental techniques and in-vivo model systems. Recently gene targeting (knockout mice) and transgenic techniques have been employed to create strains of mice whose phenotypes represent human thrombotic disorders and disease states. In conjunction with this, murine coagulation models are routinely used in the process of new drug development. During the course of developing new pro-coagulant, anti-coagulant and clot dissolving drugs, basic R&D studies as well as safety, efficacy, and pharmacokinetic studies are required. Because both normal and genetically altered mice are available, the murine models are ideal for this application. Based on the fact that murine models are in prevalent use and there exist a limited number of reagents available to assist the researcher, it is imperative to develop a commercial group of reagents for use in this field of study. This project was designed to enhance the utility of mouse coagulation models by generating a battery of reagents and analytical tools comparable to what is available for application to the human system. The long-term objective is the production of a marketable line of murine derived reagents that include: a) purified murine coagulation factors for plasma reconstitution experiments and assay standardization; b) antibodies to murine coagulation factors for Western blot, immunohistochemistry, immunoprecipitation, flow cytometry and ELISA applications; and, c) ELISA assays for the zymogens, enzymes, cofactors, inhibitors, activation peptides and enzyme/inhibitor complexes of coagulation. Utilizing conventional laboratory techniques the investigators plan to purify a wide array of hemostatic proteins, generate antibodies, both polyclonal and monoclonal to these antigens, and identify and develop clinically important immunoassays with these reagents.