The Unicamp Medicinal Chemistry Center (CQMED) and the Biotech Hilab, following the Embrapii model (Brazilian Company for Industrial Research and Innovation), perfected a protocol for developing enzymes and scaling them up to be used in molecular diagnostic tests for SARS-CoV-2.
The new platform generates enzymes that meet strict quality controls and remain active for at least 60 days, even when stored at room temperature (approximately 25°C). This characteristic is particularly important for a country the size of Brazil, as, in addition to boosting national autonomy, it favors the distribution of tests to the most remote locations, where refrigeration can be a limiting factor for the conservation of the input. O study was published on the 13th in the magazine Experimental Biology and Medicine.
The enzymes produced will be used in molecular tests developed by Hilab and operated on devices point of care also produced by the company Hilab. The Hilab Molecular is a portable device that performs tests to detect genetic material, quickly and remotely. The healthcare professional places a small amount of the collected material in the reading cartridges that contain the reagent mixture, and the device reads the result of the reaction in real time. Once connected to the internet, the device sends the information for specialized analysis to Hilab's central clinical laboratory, where a specialist analyzes it and sends the report to the requester's cell phone. The result can be received by the patient within one hour.
Quality control and enzyme stability
The great difference of the new protocol was the optimization of planning and the enzyme purification process, to increase production efficiency, reducing human manipulation and, therefore, possible contamination. “We developed a quality control platform that involves a check list of extra rehearsals. This rigorous control is the same as that carried out on commercial and imported products”, explains Lucas de Souza, author of the study and post-doctoral researcher at CQMED.
Another positive feature was the long-lasting stability of the enzymes, even under adverse conditions. “Under appropriate storage conditions, that is, -20°C, enzymes are stable for at least one year. And, at temperatures of 4°C and 25°C, they remain functional for at least 60 days. This is a crucial factor for the practical use of the device, especially in remote regions with limited access to refrigeration”, notes de Souza.
The enzymes were developed for use in the detection test for genetic material from RNA viruses in the RT-LAMP modality, which is based on the use of enzymes that allow amplification of the material under isothermal conditions. The technique began to be more explored during the covid-19 pandemic. “RT-LAMP does not require expensive equipment, unlike RT-PCR, which requires expensive and difficult-to-handle equipment”, explains Ítalo Esposti, author of the study and doctoral student at CQMED. Another advantage of RT-LAMP is that samples do not need to receive initial treatment as in RT-PCR. “In RT-LAMP, we can use 'raw samples' such as swab nasal, saliva, so we eliminated the step of extracting RNA from samples, necessary for RT-PCR, which can be complex and time-consuming”, adds Esposti.
Hilab's device was developed for the LAMP technique (Loop-Mediated Isothermal Amplification). The device is a small thermocycler, weighing approximately 200g, capable of changing temperatures quickly, in addition to having an RGB sensor. “As the reaction changes color, this means that the sample is positive for the presence of the virus. The device captures images over time to generate a graph that is interpreted by artificial intelligence. A Hilab specialist does a double check to ensure that the result is correct”, explains Bruna Raddatz, author of the study and coordinator of Hilab's molecular biology research and development team.
National autonomy and innovative public health solutions
Enzymes used to detect microorganisms in diagnostic tests are typically imported from the United States or Europe and require careful transportation, given the sensitivity of the reagent. “These imported enzymes take time to arrive and can represent approximately 75% of the final value of the test. So, the goal of the partnership with CQMED was to develop enzymes nationally as a strategy for autonomy, lower costs and access for the population to modern molecular tests”, explains Raddatz.
The new enzymes are already being used by Hilab's molecular biology team. “We are currently adapting the same system to detect a variety of viruses and bacteria for use in remote areas, hospitals and blood banks”, explains Katlin Massirer, study author and coordinator of CQMED, an accredited unit of Embrapii.
“As the world continues to face infectious disease threats, the development of modern, reliable and versatile solutions plays a vital role in protecting public health and expanding more accessible and comprehensive diagnostic capabilities”, comments Bernardo Almeida, director doctor at Hilab and author of the study. The next tests that will be incorporated into the platform will be to identify HPV, arboviruses and respiratory viruses.
“These transformative advances in public health in the country are being highly driven by the interaction between universities and companies with the support of the industrial innovation association Embrapii”, concludes Massirer.
To know more:
Lucas Rodrigo de Souza, Italo Esposti Poly da Silva, Gabriele Celis-Silva, Bruna Winkert Raddatz, Louise Matiê Imamura, Edson Yu Sin Kim, Gabriel Vieira Valderrama, Halanna de Paula Riedi, Sergio Renato Rogal Jr, Bernardo Montesanti Machado de Almeida, Marcus Vinícius Mazega Figueredo, Mario Henrique Bengtson and Katlin Brauer Massirer.
Access the article: Improved protocol for Bst polymerase and reverse transcriptase production and application to a point-of-care diagnostics system. Experimental Biology and Medicine. 2023;0(0). It hurts:10.1177/15353702231215815
About CQMED
The Center for Medicinal Chemistry – CQMED, located at the Center for Molecular Biology and Genetic Engineering (CBMEG), is linked to the Coordination of Centers and Nuclei (COCEN) at Unicamp, in Campinas (SP). The center specializes in the early stages of developing new drugs and molecular diagnostics. To this end, it developed a platform for discovering molecules that inhibit specific targets related to human diseases. CQMED was founded in 2015 and is currently accredited as an Embrapii Unit (Brazilian Research and Innovation Company) and is part of the National Institutes of Science and Technology Program – INCT of CNPq (National Council for Scientific and Technological Development).
About Hilab
Hilab is a Biotech 100% Brazilian, founded in Curitiba and also headquartered in Manaus, which develops devices and technologies nationally for clinical analysis. Its innovative approach allows exams to be carried out using just a few drops of blood.
Hilab adopts a disruptive model that eliminates the need to transport biological samples. This innovative approach makes it possible to deliver results in a matter of minutes, providing a more agile and efficient experience for patients. It uses an advanced technological platform that features Artificial Intelligence that assists healthcare professionals in clinical analyses, generating more accurate and safe results. This combination allows exams to be carried out anywhere in the world, creating a decentralized laboratory.
Furthermore, it revolutionizes the patient experience by offering less uncomfortable collections at your fingertips. With the use of innovative devices, Hilab simplifies the process of delivering results, which takes place in just a few minutes.
About EMBRAPII
The Brazilian Industrial Research and Innovation Company was created in 2013 as a social organization that has autonomy in managing funds from the Ministry of Science, Technology and Innovation and the Ministry of Education, with the aim of stimulating innovation in Brazilian industry, promoting interaction between institutions technological research and companies in the industrial sector.
Its operating model provides for the financing of up to one third of the total cost of each approved project, with non-refundable resources (i.e., the industry does not need to return the amount contributed), and the remainder is divided between the industry and the EMBRAPII units. .
