Protein Stability and Functionality Enhancement Method

申请人的描述：一种新颖的通用协议，'ids.PCR'（专利 待决），已经被开发用于独特地修饰基因序列，并且因此可以被 用于将新的和期望的特性进化成基因产物。Ids.PCR去 超越现有的“定向进化”技术，提供更大的多样性 然而，它可以被“调整”以实现低水平的随机化， 在需要的地方。 Ids.PCR是一种基于PCR的技术的首字母缩写，该技术促进了随机扩增。 DNA内单或多核苷酸插入、缺失和/或取代 顺序这种多方面的能力有助于创建随机 独特基因或基因片段的文库，从其编码具有 可以选择和隔离期望的特性。结合 合适的筛选，净效应是一个强大的加速进化 修改现有基因序列以产生期望的/最佳的 蛋白质特性，如稳定性和特异性。 ids.PCR的应用包括开发和优化新的 满足关键生物医学需求的产品，包括蛋白质 药物如蛋白质生长因子、抗体和疫苗， 以及用于小分子药物生产的优化酶。具体 本提案I期的研究目标是使用β-内酰胺酶作为 用于生产表现出增强的修饰酶的模型系统 功能和稳定性。 拟议的商业应用： 建议的ID PCR方法可用于开发独特的蛋白质， 生物医学应用所需的特性，包括（作为实例）： 具有增强的稳定性、改善的半衰期和减少的有害物质的蛋白质药物 副作用和毒性。B）具有增强免疫系统的安全有效的疫苗 刺激的特性。 商业生产的蛋白质药物（例如G-CSF） 还可以改进。

APPLICANT'S DESCRIPTION: A novel and versatile protocol, 'ids.PCR' (patent pending), has been developed to uniquely modify gene sequence, and can thus be used to evolve new and desirable properties into gene products. Ids.PCR goes beyond existing "directed evolution" technologies, affording greater diversity of molecules, and yet can be 'tuned' to effect low levels of randomization where desired. Ids.PCR is an acronym for a PCR-based technology that facilitates the random insertion, deletion and/or substitution of mono-or polynucleotides within a DNA sequence. This multifaceted capability facilitates the creation of random libraries of unique genes or sections of genes from which encoded proteins with desired characteristics can be selected and isolated. In conjunction with suitable screening, the net effect is a powerful accelerated evolutionary process that modifies the existing gene sequence to engender desirable/optimal protein characteristics, such as stability and specificity. Applications of ids.PCR include the development and optimization of novel products addressing critical biomedical needs, including protein pharmaceuticals such as protein growth factors, antibodies, and vaccines, as well as optimized enzymes for small molecule drug production. The specific research objective of this proposal for Phase I is to use beta-lactamase as a model system for the production of modified enzymes exhibiting enhanced function and stability. PROPOSED COMMERCIAL APPLICATIONS: The proposed ids. PCR methodology could be utilized to develop unique proteins with desired characteristics for biomedical applications including (as examples): a)Improved protein pharmaceuticals with enhanced stability, improved half-life, and reduced harmful side effects and toxicities. b) Safe and effective vaccines with enhanced immune system stimulating characteristics. Commercially produced protein pharmaceuticals (e.g. G-CSF) could be improved upon.