Protein Stability and Functionality Enhancement Method

申请人描述：一种新颖且通用的协议，id。PCR(专利

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.