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BioApatIgG - immunological diagnostic assay of the low cost and high performance

Abstract

In December 2019, the World Health Organization was informed of a number of cases of pneumonia of unknown aetiology in Wuhan, China. Several subsequent investigations identified a new coronavirus, now called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in samples of affected patients. The highly sensitive and specific laboratory diagnosis is important for the rapid control of the evolution of SARS-CoV-2 2019 associated coronavirus disease (COVID-19). In Brazil, on February 26th, the first patient with COVID-19 was diagnosed and registered. Since then the curve has grown logarithmically from new cases and the need for inputs for fast and accurate tests is a demand of high importance. Furthermore, there are recommended molecular tests for viral RNA detection in human fluids. Also, detection tests for monitoring people who have already developed immunity to the disease are of fundamental importance for monitoring the actual progress of the virus. In Brazil, the inputs for the production of both molecular tests and immunoassays are imported today, and with the advancement of the pandemic, the logistics of materials and high prices lead to the need to create local production of biotechnology inputs for use in immunoassays and molecular tests. BIOLINKER, a company specialized in rapid protein synthesis with an in vitro transcription and translation system, proposes, in partnership with Professor Ester Sabino of the Institute of Tropical Medicine (USP) and Professor Almidar Tanuri's UFRJ Virology Group, to supply ELISA reaction kits, standardized for the detection of circulating IgG against the COVID-19 virus in patient serum. Our results with the aptamers have demonstrated higher sensitivity and specificity to binding at constant fraction of IgG from humans. This results was validated and developed in the PIPE-phase 1 n. of the process 2017/22801-6. We combined it with biotin and the detection was by peroxidase combined with avidin. We want to use this tool to evaluate the detection of IgG against anti-nucleocapsid N, anti-RBS (spike receptor binding region) and viral-dependent anti-polymerase RdRp-RNA in the serum. Our objective in this promotion is to establish an antigen production scale that initially produced by cell-free protein synthesis and then was immobilized in polystyrene plates at 85 % of purity. We will manufacture the buffers and aptamer anti-IgG conjugated with biotin for detection in serum of patients in screening provided by the Institute of Tropical Medicine - FM-USP, enabling a fast and less expensive trial for epidemiological screening of the disease. Our estimated costs are low, since the cost of production of aptamers is relatively low, as well as the production of the antigen via cell-free, since the company has its own plasmids of expression and protocols already established. (AU)

Articles published in Pesquisa para Inovação FAPESP about research grant:
Researchers develop 10-minute COVID-19 diagnostic test that wage workers can afford 
Articles published in Agência FAPESP Newsletter about the research grant:
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