Lab-on-a-chip integrated protein synthesis: development of DNA aptamers to be used for purification steps

• Grant number: 17/22801-6
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Duration: February 01, 2019 - December 31, 2019
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Mona das Neves Oliveira
• Grantee: Mona das Neves Oliveira
• Host Company: Biolinker Biologia Sintética Eireli
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais
• City: São Paulo
• Associated grant(s): 20/05023-2 - BioApatIgG - immunological diagnostic assay of the low cost and high performance, AP.PIPE
• Associated scholarship(s): 19/05030-1 - Lab-on-a-chip integrated protein synthesis: development of DNA aptamers to be use for purification steps, BP.TT; 19/07173-4 - Lab-on-a-chip integrated protein synthesis: development of DNA aptamers to be use for purification steps, BP.PIPE

Abstract

In this project, we aim to build a miniaturized, integrated system with parts to be developed in different institutes and to be marketed by the company Biolinker Synthetic Biology. Lab-on-Chip integrated protein synthesis was born as an ambitious project that aims to raise the quality of research, facilitating the work and reducing steps in the production of proteins for research laboratories in the world. Cell free protein synthesis (CFPS) is a technique that has been used in research laboratories due to flexibility and reduction of the time spent in the protein production both in basic and applied research. Technological advances and modern application of biotechnology to obtain and modelling of new chemical structures allowed by CFPS are of a great magnitude. This new technology allows the addition of non-canonical amino acids, as well as the biosynthesis of toxins, which were limitations in the process based on transfection and expression via cells (so called cell-based protein synthesis). Additionally, this research field is quite beneficial for the pharmacological context and tends to impact concepts such as chemistry and bioconjugation therapy. (Zemella, Thoring, Hoffmeister, & Kubick, 2015).In this context, Biolinker aims to create a model that optimizes production and biosynthesis of proteins from eukaryotic and prokaryotic cells. The model proposes to integrate in a single system the reactions of transcription and translation, followed by the purification with detection and quantification of the desired protein. In partnership with Prof. Dr. Henning Ulrich of the Chemical Institute of USP, we aim to develop aptamers to be used in the purification step and set in microchip purification using the SELEX technique (Systematic Evolution of Ligands by Exponential enrichment, Szostak) (Tuerk and Gold, 1990; Ellington and Szostak 1990); The major advantages of the use of aptamers biotechnology range from the possibility of replacing antibodies, independent acquisition of animal use and the possibility to be easily modified for their respective applications. Commercially, it is more advantageous to build more competitive products with use of input of relatively low value and high versatility of use. Due to the small size of aptamers, they can be effectively fixed with high densities, which is also an important factor in building and increasing the connection sensitivity in bioanalytical machines (Iliuk et al 2011). Finally, the company plans to use the technology "Lab-On-Chip" to be developed through partnerships with Prof. Gianluca Grenci, Senior Scientist at the Mechanobiology Institute, NUS Singapore and the headquarters of Biolinker in Slovenia-NIB. For machine supplement that is being developed, we aim to partner with the research group of the Fraunhofer Institute for Cell Therapy and Immunology IZI, Germany led by Prof. Stefan Kubick, Head of Department "Cell-free and Cell-based Bioproduction" which will be responsible for the development and production of kits to be used in microchips. (AU)