Production of scFv, Fab, and IgG of CR3022 Antibodies Against SARS-CoV-2 Using Silkworm-Baculovirus Expression System

Ebihara, Takeru; Masuda, Akihiro; Takahashi, Daisuke; Hino, Masato; Mon, Hiroaki; Kakino, Kohei; Fujii, Tsuguru; Fujita, Ryosuke; Ueda, Tadashi; Lee, Jae Man; Kusakabe, Takahiro

doi:10.1007/s12033-021-00373-0

Cited by 9 publications

(7 citation statements)

References 51 publications

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“…F(ab') 2 are produced through the enzymatic cleavage of immunoglobulins and conserve both the antigenbinding region and part of the hinge region. The use of monoclonal F(ab') 2 for the treatment of SARS-CoV-2 has already shown efficacious potential in various studies utilizing different methods to raise the particle (8)(9)(10). However, one important caveat of monoclonal antibodies that extends to most current F(ab') 2 is that they are designed to target one specific epitope, rendering treatment inefficacious in the event of mutation.…”

Section: Introductionmentioning

confidence: 99%

Passive immunization with equine RBD-specific Fab protects K18-hACE2-mice against Alpha or Beta variants of SARS-CoV-2

et al. 2022

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Emergence of variants of concern (VOC) during the COVID-19 pandemic has contributed to the decreased efficacy of therapeutic monoclonal antibody treatments for severe cases of SARS-CoV-2 infection. In addition, the cost of creating these therapeutic treatments is high, making their implementation in low- to middle-income countries devastated by the pandemic very difficult. Here, we explored the use of polyclonal EpF(ab’)2 antibodies generated through the immunization of horses with SARS-CoV-2 WA-1 RBD conjugated to HBsAg nanoparticles as a low-cost therapeutic treatment for severe cases of disease. We determined that the equine EpF(ab’)2 bind RBD and neutralize ACE2 receptor binding by virus for all VOC strains tested except Omicron. Despite its relatively quick clearance from peripheral circulation, a 100μg dose of EpF(ab’)2 was able to fully protect mice against severe disease phenotypes following intranasal SARS-CoV-2 challenge with Alpha and Beta variants. EpF(ab’)2 administration increased survival while subsequently lowering disease scores and viral RNA burden in disease-relevant tissues. No significant improvement in survival outcomes or disease scores was observed in EpF(ab’)2-treated mice challenged using the Delta variant at 10μg or 100µg doses. Overall, the data presented here provide a proof of concept for the use of EpF(ab’)2 in the prevention of severe SARS-CoV-2 infections and underscore the need for either variant-specific treatments or variant-independent therapeutics for COVID-19.

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Section: Introductionmentioning

confidence: 99%

Passive immunization with equine RBD-specific Fab protects K18-hACE2-mice against Alpha or Beta variants of SARS-CoV-2

et al. 2022

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“…Codon optimization. High Five/ExpiSf9/silkworm larvae Affinity chromatography scFv, Fab, IgY and IgG antibody productions [ 94 , 95 , 97 , 98 ] Spike protein and receptor binding domain Signal peptide and peptide tag added. Biotinylation.…”

Section: Application Of Baculovirus Expression Vector System (Bev) Fo...mentioning

confidence: 99%

“…A monoclonal antibody against RBD, CR3022, has been produced using a silkworm-baculovirus expression vector system in three formats (scFv, Fab, and IgG) [ 95 ]. The affinity of the antibody towards S protein was equivalent to the one that was produced in the mammalian expression system.…”

Section: Application Of Baculovirus Expression Vector System (Bev) Fo...mentioning

confidence: 99%

Application of Baculovirus Expression Vector system (BEV) for COVID-19 diagnostics and therapeutics: a review

Azali

Mohamed

Harun

et al. 2022

Journal of Genetic Engineering and Biotechnology

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Background The baculovirus expression vector system has been developed for expressing a wide range of proteins, including enzymes, glycoproteins, recombinant viruses, and vaccines. The availability of the SARS-CoV-2 genome sequence has enabled the synthesis of SARS-CoV2 proteins in a baculovirus-insect cell platform for various applications. Main body of the abstract The most cloned SARS-CoV-2 protein is the spike protein, which plays a critical role in SARS-CoV-2 infection. It is available in its whole length or as subunits like S1 or the receptor-binding domain (RBD). Non-structural proteins (Nsps), another recombinant SARS-CoV-2 protein generated by the baculovirus expression vector system (BEV), are used in the identification of new medications or the repurposing of existing therapies for the treatment of COVID-19. Non-SARS-CoV-2 proteins generated by BEV for SARS-CoV-2 diagnosis or treatment include moloney murine leukemia virus reverse transcriptase (MMLVRT), angiotensin converting enzyme 2 (ACE2), therapeutic proteins, and recombinant antibodies. The recombinant proteins were modified to boost the yield or to stabilize the protein. Conclusion This review covers the wide application of the recombinant protein produced using the baculovirus expression technology for COVID-19 research. A lot of improvements have been made to produce functional proteins with high yields. However, there is still room for improvement and there are parts of this field of research that have not been investigated yet.

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“…Therefore, by evaluating the neutralizing activity of antibodies, SPR assays proved their utility to target viral surface proteins and inhibit SARS-CoV-2 spike interaction with its receptor. The binding of scFv, Fab, and IgG (immunoglobulin G) of CR3022 antibodies to the RBD of SARS-CoV-2 has also been investigated by SPR in a silkworm-baculovirus expression system [56]. SPR analysis demonstrated that the IgG antibodies produced by silkworms had almost the same affinity to S protein that those produced in mammalian expression system.…”

Section: Spr Biosensors As Diagnostic Tools To Detect Neutralizing Antibodiesmentioning

confidence: 99%

Current Trends in SPR Biosensing of SARS-CoV-2 Entry Inhibitors

Mauriz

Lechuga

2021

Chemosensors

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The emerging risk of viral diseases has triggered the search for preventive and therapeutic agents. Since the beginning of the COVID-19 pandemic, greater efforts have been devoted to investigating virus entry mechanisms into host cells. The feasibility of plasmonic sensing technologies for screening interactions of small molecules in real time, while providing the pharmacokinetic drug profiling of potential antiviral compounds, offers an advantageous approach over other biophysical methods. This review summarizes recent advancements in the drug discovery process of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) inhibitors using Surface Plasmon Resonance (SPR) biosensors. A variety of SPR assay formats are discussed according to the binding kinetics and drug efficacies of both natural products and repurposed drugs. Special attention has been given to the targeting of antiviral agents that block the receptor binding domain of the spike protein (RBD-S) and the main protease (3CLpro) of SARS-CoV-2. The functionality of plasmonic biosensors for high-throughput screening of entry virus inhibitors was also reviewed taking into account experimental parameters (binding affinities, selectivity, stability), potential limitations and future applications.

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Production of scFv, Fab, and IgG of CR3022 Antibodies Against SARS-CoV-2 Using Silkworm-Baculovirus Expression System

Cited by 9 publications

References 51 publications

Passive immunization with equine RBD-specific Fab protects K18-hACE2-mice against Alpha or Beta variants of SARS-CoV-2

Passive immunization with equine RBD-specific Fab protects K18-hACE2-mice against Alpha or Beta variants of SARS-CoV-2

Application of Baculovirus Expression Vector system (BEV) for COVID-19 diagnostics and therapeutics: a review

Current Trends in SPR Biosensing of SARS-CoV-2 Entry Inhibitors

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