Posters
Academic/Professional Position (Other)
PhD
Presentation Type
Poster
Discipline Track
Community/Public Health
Abstract Type
Research/Clinical
Abstract
Background: COVID-19 represents a significant threat to global human health. SARS-CoV-2, the etiologic viral agent, needs to be under-covered at the structural biology level to facilitate the rational design of diagnostic tests and vaccine candidates. SARS-CoV-2 Receptor Binding Domain of Spike protein (RBD-S) acts as the key to open the gate, to enter the cells during infection. Thus, it is a stronger candidate for designing effective antigens for vaccines and diagnostics. Here, we relied on the viral DNA codifying to RBD-S to use synthetic biology for optimizing the recombinant expression of this (rRBD-S) as a proof of concept of rational designs of bioprocess for vaccine candidates and immunogens to improved rapid diagnostic tests.
Methods: rRBD-S coding sequences inspired on RBD-S ectodomain from SARS-CoV-2 were designed, codon-optimized, tagged, synthesized, cloned in an expression vector (pD444-MR), and transformed into C41(DE3)pLysS E. coli strain. Expression of recombinant RBD-S was resulting in a protein purified using Ni-IMAC (Nickel Immobilized metal affinity chromatography).
Results: rRBD-S produced result in a ~30KDa protein with yields of 4.618 gr L-1. Protein was recovered from the bacterial soluble fraction without refolding process.
Conclusions: rRBD-S is an important tool for immunity diagnostics as Lateral-Flow-Devices, structural biology studies, and even as vaccine candidate for combating SARS-CoV-2. Notably considering the advantages of rational subunit vaccines for immune response against other vaccines technologies whose effectiveness in the long-term process has not been demonstrated yet.
Acknowledgements: We thank HIDALGO´s Council of Science, Technology and Innovation (CITNOVA) for the GRANT 20201122 to LMRM.
Recommended Citation
Rodriguez, Luis Mario; Rodriguez, Mario Alberto; Barrera-Saldaña, Hugo A.; Martinez, Rocio Marisol; and Rodriguez, Daniela, "Rational design for the recombinant expression of the Receptor Binding Domain of SARS-CoV-2 Spike Glycoprotein" (2023). Research Symposium. 86.
https://scholarworks.utrgv.edu/somrs/theme1/posters/86
Included in
Rational design for the recombinant expression of the Receptor Binding Domain of SARS-CoV-2 Spike Glycoprotein
Background: COVID-19 represents a significant threat to global human health. SARS-CoV-2, the etiologic viral agent, needs to be under-covered at the structural biology level to facilitate the rational design of diagnostic tests and vaccine candidates. SARS-CoV-2 Receptor Binding Domain of Spike protein (RBD-S) acts as the key to open the gate, to enter the cells during infection. Thus, it is a stronger candidate for designing effective antigens for vaccines and diagnostics. Here, we relied on the viral DNA codifying to RBD-S to use synthetic biology for optimizing the recombinant expression of this (rRBD-S) as a proof of concept of rational designs of bioprocess for vaccine candidates and immunogens to improved rapid diagnostic tests.
Methods: rRBD-S coding sequences inspired on RBD-S ectodomain from SARS-CoV-2 were designed, codon-optimized, tagged, synthesized, cloned in an expression vector (pD444-MR), and transformed into C41(DE3)pLysS E. coli strain. Expression of recombinant RBD-S was resulting in a protein purified using Ni-IMAC (Nickel Immobilized metal affinity chromatography).
Results: rRBD-S produced result in a ~30KDa protein with yields of 4.618 gr L-1. Protein was recovered from the bacterial soluble fraction without refolding process.
Conclusions: rRBD-S is an important tool for immunity diagnostics as Lateral-Flow-Devices, structural biology studies, and even as vaccine candidate for combating SARS-CoV-2. Notably considering the advantages of rational subunit vaccines for immune response against other vaccines technologies whose effectiveness in the long-term process has not been demonstrated yet.
Acknowledgements: We thank HIDALGO´s Council of Science, Technology and Innovation (CITNOVA) for the GRANT 20201122 to LMRM.