Development of a in-house hot start PCR kit to improve national research & development

• Grant number: 19/09113-9
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Duration: February 01, 2020 - December 31, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Bruna Tatsue Grichoswski Nakagawa
• Grantee: Bruna Tatsue Grichoswski Nakagawa
• Host Company: Ecra Biotec Serviços e Pesquisas Ltda
• CNAE: Serviços combinados de escritório e apoio administrativo
• City: Santo André
• Associated researchers: Fabio Trigo Raya ; Robson Tramontina
• Associated scholarship(s): 20/06377-2 - Development of a in-house hot start PCR kit to improve national research & development, BP.PIPE

Abstract

Molecular diagnosis assays conducted in national governmental laboratories (Instituto Evandro Chagas, Lacen, Fundação Oswaldo Cruz among others) or in research centers, require the availability of a DNA polymerase enzyme. DNA polymerase allows the in vitro replication of the DNA under optimal conditions. Temperature is a factor that exert a high impact on the enzyme activity. Although the optimal temperature is 95ºC, DNA polymerase is also able to polymerize at lower temperatures (below 50C), causing limitations during mixture preparation. For that reason, DNA polymerases with a hot start trigger were developed. These enzyme are inactivated by the presence of an antibody, aptamers, Nano molecule, etc. and when temperature increases in the beginning of a PCR reaction, the inhibitory element is released and the reaction proceeds normally. In the Brazilian context, the access to this enzyme is limited by the cost (overpriced when imported) and the time span required for the overseas shipment. This scenario is particularly concerning when diagnosis kits to detect Zika or Dengue virus (for example), depend upon the availability of this enzyme. To face that limitation, ECRABIOTEC, proposes to develop an in-house, high fidelity hot start DNA polymerase kit fundamental to support the development of the detections kits for aforementioned diseases. Since the R&D will be 100% Brazilian, the final cost and the shipment will be significantly more attractive to researchers. Group efforts have allowed the development of a high fidelity DNA polymerase, and the prototype was made available to 10 different coworkers in the research area, to test and give their feedback. The outcome was positive and therefore ECRABIOTEC research team, decided to submit a proposal to PIPE fase 1, to use the in-house developed DNA polymerase and test the technical viability of adding the hot start mechanism. A strategy using aptamers and primers will be tested and the best one will be submitted to the PIPE fase 2 to implement the idea and scale up the process. The preliminary results obtained so far, attracted the attention of the business players in the pharmaceutical context, which lead to the beginning of ECRABIOTEC, which came to substitute ECRA healthcare, a 3 years old company with expertise in the market. The submission of this project aims to start a new research line in the biotechnology field, by developing a national DNA polymerase kit and accelerating the Brazilian research competences. (AU)