Protein Stability and Functionality Enhancement Method

蛋白质稳定性和功能增强方法

• 批准号: 6321910
• 负责人: Eric Lietz
• 金额: $ 9.93万
• 依托单位: GENOPSYS, INC.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6321910
• 关键词: beta lactamase; biochemical evolution; biotechnology; chemical registry /resource; chemical stability; combinatorial chemistry; enzyme activity; evaluation /testing; gene mutation; genetic manipulation; method development; mutant; nucleic acid chemical synthesis; nucleic acid sequence; polymerase chain reaction; protein structure function; recombinant proteins

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.

申请人描述：一种新颖而通用的方案，‘ids.PCR’(专利 待定)，已经被开发成唯一地修改基因序列，因此可以 用于将新的和理想的特性进化成基因产品。Ids.PCR走了 超越现有的“定向进化”技术，提供更大的多样性 分子的数量，而且还可以进行调整，以实现低水平的随机化 在想要的地方。 Ids.PCR是基于PCR的技术的首字母缩写，该技术促进了随机 DNA中单核苷酸或多核苷酸的插入、缺失和/或替换 序列。这种多方面的能力有助于创建随机 用来编码蛋白质的独特基因库或基因片段 可以选择和隔离所需的特性。与 合适的筛选，净效果是一个强大的加速进化 修改现有基因序列以产生所需的/最佳的过程 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型的 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，如 以及用于小分子药物生产的优化酶。具体的 该方案第一阶段的研究目标是将β-内酰胺酶用作 一种具有增强型修饰酶生产模型系统 功能和稳定性。 建议的商业应用： 建议的身份证件。可以利用聚合酶链式反应方法来开发独特的蛋白质 生物医学应用所需的特性，包括(例如)：a)改进 增强稳定性、改善半衰期、减少有害物质的蛋白质药物 副作用和毒性。B)增强免疫系统的安全有效疫苗 激动人心的特点。商业化生产的蛋白质药物(如G-CSF) 可能会有所改进。