Luis Giovani de Oliveira Guardalini

@butantan.gov.br

Laboratório de Biotecnologia Viral
Instituto Butantan

Luis Giovani de Oliveira Guardalini


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https://researchid.co/giovani_guardalini
8

Scopus Publications

147

Scholar Citations

8

Scholar h-index

7

Scholar i10-index

Scopus Publications

  • Non-clinical analysis of virus-like particles (VLP) containing SARS-CoV-2 vaccine antigens
    Luis Giovani Oliveira Guardalini, Isabela Mancini Martins, Thaissa Consoni Bernardino, Wagner Quintilio, Kaio Moraes de Farias, Soraia Attie Calil Jorge, Durvanei Augusto Maria
    Vaccine, 2026
  • Controlled vs. non-controlled culture systems for SARS-CoV-2 VLP production using the baculovirus/SF9 platform
    Luis Giovani de Oliveira Guardalini, Felipe Moura Dias, Thaissa Consoni Bernardino, Jaci Leme, Henrique Coelho de Oliveira, Aldo Tonso, Eutimio Gustavo Fernández-Núñez, Soraia Attie Calil Jorge
    Preparative Biochemistry and Biotechnology, 2026
    COVID-19 is an infectious disease resulting from the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which emerged as a global pandemic in 2020. Virus-like particles (VLP) are artificially generated nanoparticles composed of a subset of viral components that closely mimic the native virus in size, structure, and surface composition. These features contribute to their robust immunogenic potential, stimulating both antibody-mediated and cell-mediated immune responses. This study aimed to evaluate the expression of SARS-CoV-2 structural proteins and the VLP production using a baculovirus-insect cell system. Two monocistronic recombinant baculoviruses, each carrying either the spike (S) or nucleocapsid (N) gene of SARS-CoV-2, were employed in two different culture systems: Schott flasks and a stirred-tank bioreactor. The Sf9 cells showed substantial differences between the Schott flask and stirred tank bioreactor in terms of metabolism, virus titer, and cell death after infection. Both proteins' expressions were confirmed in the two culture systems assessed. However, VLP formation (69.1-78.2 nm) was only confirmed in the co-infection setup with both monocistronic recombinant baculoviruses. In contrast, infection with only the S-protein baculovirus failed to produce VLP in our findings.
  • Monitoring by Raman spectroscopy of rabies virus-like particles production since the initial development stages
    Luis Giovani Oliveira Guardalini, Rafaela Morais Rangel, Jaci Leme, Thaissa Consoni Bernardino, Suellen Regina da Silveira, Aldo Tonso, Soraia Attie Calil Jorge, Eutimio Gustavo Fernández Núñez
    Journal of Chemical Technology and Biotechnology, 2024
    BACKGROUNDThis study aimed to establish chemometric models using Raman spectroscopy data for biochemical monitoring of rabies Virus‐Like Particles (VLP) production based on baculovirus/insect cell system. The models were developed using fresh and stored samples from the initial development stages (Schott culture flasks). The following modeling techniques were assessed: partial least squares (PLS) and artificial neural networks (ANN). The effects of spectral filtering approaches, spectral ranges (400–1850 cm−1; 100–3425 cm−1), and sample cryopreservation were also considered. The applicability of the models was evaluated using experimental data from assays carried out in a benchtop bioreactor.RESULTSThe results showed that the prediction capacity of the chemometric models was negatively impacted when samples from rabies VLP production were cryopreserved. Further studies are needed to confirm the maximum storage time for samples (< 4 months) without a significant difference in model predictions compared to those from an in line database. The dilution of the sample should be kept constant throughout the rabies VLP development stages. A nonlinear correlation was observed between dilution and the predicted values of biochemical parameters from Raman spectral data. The choice of spectral filtering has a major impact on the prediction accuracy of chemometric models.CONCLUSIONThe optimal filtering approach should be individually optimized for each biochemical parameter. The ANN models were significantly more suitable for biochemical monitoring than the PLS approach. The 400–1850 cm−1 Raman shift range is recommended for biochemical monitoring of rabies VLP using a baculovirus/insect cell platform when samples are cell‐free. © 2023 Society of Chemical Industry (SCI).
  • Sf9 Cell Metabolism Throughout the Recombinant Baculovirus and Rabies Virus-Like Particles Production in Two Culture Systems
    Luis Giovani Oliveira Guardalini, Jaci Leme, Paulo Eduardo da Silva Cavalcante, Renata Gois de Mello, Thaissa Consoni Bernardino, Simone Gonçalves Silva Jared, Marta Maria Antoniazzi, Renato Mancini Astray, Aldo Tonso, Eutimio Gustavo Fernández Núñez, Soraia Attie Calil Jorge
    Molecular Biotechnology, 2024
  • Sf9 Cells Metabolism and Viability When Coinfected with Two Monocistronic Baculoviruses to Produce Rabies Virus-like Particles
    Jaci Leme, Luis Giovani Oliveira Guardalini, Thaissa Consoni Bernardino, Renato Mancini Astray, Aldo Tonso, Eutimio Gustavo Fernández Núñez, Soraia Attie Calil Jorge
    Molecular Biotechnology, 2023
  • Zika virus-like particles (VLPs) produced in insect cells
    Renata Gois de Mello, Thaissa Consoni Bernardino, Luis Giovani Oliveira Guardalini, Renato Mancini Astray, Marta Maria Antoniazzi, Simone Gonçalves Silva Jared, Eutimio Gustavo Fernández Núñez, Soraia Attie Calil Jorge
    Frontiers in Pharmacology, 2023
    Introdutcion: The Zika virus (ZIKV) infections are a healthcare concern mostly in the Americas, Africa, and Asia but have increased its endemicity area beyond these geographical regions. Due to the advances in infections by Zika virus, it is imperative to develop diagnostic and preventive tools against this viral agent. Virus-like particles (VLPs) appear as a suitable approach for use as antiviral vaccines.Methods: In this work, a methodology was established to produce virus-like particles containing the structural proteins, C, prM, and E of Zika virus produced in insect cells using the gene expression system derived from baculovirus. The vector pFast- CprME -ZIKV was constructed containing the gene sequences of Zika virus structural proteins and it was used to generate the recombinant bacmids (Bac- CprME -ZIKV) through transformation into DH10BacTM cells. The Bac- CprME -ZIKV was transfected in Spodoptera frugiperda (Sf9) insect cells and batches of BV- CprME -ZIKV were obtained by infection assays using a multiplicity of infection of 2. The Sf9 cells were infected, and the supernatant was collected 96 h post-infection. The expression of the CprME -ZIKV protein on the cell surface could be observed by immunochemical assays. To concentrate and purify virus-like particles, the sucrose and iodixanol gradients were evaluated, and the correct CprME -ZIKV proteins’ conformation was evaluated by the Western blot assay. The virus-like particles were also analyzed and characterized by transmission electron microscopy.Results and discussion: Spherical structures like the native Zika virus from 50 to 65 nm containing the CprME -ZIKV proteins on their surface were observed in micrographs. The results obtained can be useful in the development path for a vaccine candidate against Zika virus.
  • Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms
    Luis Giovani Oliveira Guardalini, Paulo Eduardo da Silva Cavalcante, Jaci Leme, Renata Gois de Mello, Thaissa Consoni Bernardino, Simone Gonçalves Silva Jared, Marta Maria Antoniazzi, Renato Mancini Astray, Aldo Tonso, Eutimio Gustavo Fernández Núñez, Soraia Attie Calil Jorge
    Vaccines, 2023
    This work aimed to assess, following upstream optimization in Schott flasks, the scalability from this culture platform to a stirred-tank bioreactor in order to yield rabies-recombinant baculovirus, bearing genes of G (BVG) and M (BVM) proteins, and to obtain rabies virus-like particles (VLP) from them, using Sf9 insect cells as a host. Equivalent assays in Schott flasks and a bioreactor were performed to compare both systems and a multivariate statistical approach was also carried out to maximize VLP production as a function of BVG and BVM’s multiplicity of infection (MOI) and harvest time (HT). Viable cell density, cell viability, virus titer, BVG and BVM quantification by dot-blot, and BVG quantification by Enzyme-Linked Immunosorbent Assay (ELISA) were monitored throughout the assays. Furthermore, transmission electron microscopy was used to characterize rabies VLP. The optimal combination for maximum VLP expression was BVG and BVM MOI of 2.3 pfu/cell and 5.1 pfu/cell, respectively, and 108 h of harvest time. The current study confirmed that the utilization of Schott flasks and a benchtop bioreactor under the conditions applied herein are equivalent regarding the cell death kinetics corresponding to the recombinant baculovirus infection process in Sf9 cells. According to the results, the hydrodynamic and chemical differences in both systems seem to greatly affect the virus and VLP integrity after release.
  • Evaluation of the applicability and effectiveness of a molecular strategy for identifying weak D and DEL phenotype among D– blood donors of mixed origin exhibiting high frequency of RHD*Ψ
    Marcia Regina Dezan, Luis Giovani O. Guardalini, Elaine Pessoa, Ingrid Helena Ribeiro, Valeria Brito Oliveira, Fabio Luz, Denise Rossite Novac, António Gallucci, Silvia Bonifácio, Francisco Gomes, José E. Levi, Alexandre C. Pereira, Jose E. Krieger, Alfredo Mendrone-Junior, Vanderson Rocha, Carla Luana Dinardo
    Transfusion, 2018
    Molecular tests designed to detect the presence of active RHD gene among D– donors have been successfully applied in people of European ancestry, but not in admixed populations with a considerable frequency of RHD*Ψ. Our goal was to evaluate the performance of a molecular screening tool for identifying active RHD alleles among Brazilian blood donors classified as D– C+ and/or E+.

RECENT SCHOLAR PUBLICATIONS

  • Controlled vs. non-controlled culture systems for SARS-CoV-2 VLP production using the baculovirus/SF9 platform
    LGO Guardalini, FM Dias, TC Bernardino, J Leme, HC de Oliveira, ...
    Preparative Biochemistry & Biotechnology, 1-12 , 2026
    2026
  • Non-clinical analysis of virus-like particles (VLP) containing SARS-CoV-2 vaccine antigens
    LGO Guardalini, IM Martins, TC Bernardino, W Quintilio, KM de Farias, ...
    Vaccine 79, 128406 , 2026
    2026
  • Media-dependent performance of rabies virus-like particle production under low oxygen tension in baculovirus-infected Sf9 cells
    JP Rabello, MM Teruya, FÂC Barrence, TC Bernardino, J Leme, ...
    Journal of Chemical Technology and Biotechnology, 01-10 , 2026
    2026
  • Media‐dependent performance of rabies virus‐like particle production under low oxygen tension in baculovirus‐infected Sf9 cells
    J Públio Rabello, MM Teruya, FA Correia Barrence, ...
    Journal of Chemical Technology & Biotechnology , 2026
    2026
  • Zika virus-like particle production in a stirred tank bioreactor using a baculovirus/insect cell system
    T Consoni Bernardino, MM Teruya, PE da Silva Cavalcante, ...
    Bioprocess and Biosystems Engineering 48 (12), 2137-2152 , 2025
    2025
  • Scalable approach for coronavirus-like particles making based on the spike protein using Sf9 cells
    LG de Oliveira Guardalini, TC Bernardino, FM Dias, J Leme, ...
    Protein Expression and Purification 235, 106767 , 2025
    2025
    Citations: 1
  • MIR spectroscopy in ATR mode for Sf9 insect cell line biochemical monitoring in bioprocesses
    LG de Oliveira Guardalini, GM da Trindade, J Leme, TC Bernardino, ...
    Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 338, 126174 , 2025
    2025
    Citations: 1
  • Offline Raman monitoring of SARS-CoV-2 VLP upstream stage using two culture media
    J Públio Rabello, I de Sousa Fontes, F Moura Dias, S Omae Camalhonte, ...
    Spectroscopy Letters, 1-13 , 2025
    2025
    Citations: 1
  • Multiplicity of infection and culture medium on the SARS-CoV-2 virus like-particles production by baculovirus/insect system
    LG de Oliveira Guardalini, F Moura Dias, S Omae Camalhonte, J Leme, ...
    Biotechnology Letters 47 (2), 32 , 2025
    2025
    Citations: 4
  • Chemometrics and analytical blank on the at-line monitoring of Zika-VLP production using near-infrared spectroscopy
    JP Rabello, PE da Silva Cavalcante, J Leme, VAT Dias, FAC Barrence, ...
    Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 326, 125217 , 2025
    2025
    Citations: 5
  • Inline Raman spectroscopy as process analytical technology for SARS-CoV-2 VLP production
    F Moura Dias, MM Teruya, S Omae Camalhonte, V Aragão Tejo Dias, ...
    Bioprocess and Biosystems Engineering 48 (1), 63-84 , 2025
    2025
    Citations: 4
  • Biochemical monitoring of SARS-CoV-2 VLP production stages using MIR spectroscopy and chemometrics
    FM Dias, TC Bernardino, LG de Oliveira Guardalini, JL Tanaka, J Leme, ...
    Process Biochemistry 147, 418-432 , 2024
    2024
    Citations: 3
  • Raman laser intensity and sample clarification on biochemical monitoring over Zika-VLP upstream stages
    PE da Silva Cavalcante, JP Rabello, J Leme, VAT Dias, FAC Barrence, ...
    Biochemical and Biophysical Research Communications 733, 150671 , 2024
    2024
    Citations: 3
  • Different modeling approaches for inline biochemical monitoring over the VLP-making upstream stages using Raman spectroscopy
    VAT Dias, LGO Guardalini, J Leme, TC Bernardino, SR da Silveira, ...
    Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 320, 124638 , 2024
    2024
    Citations: 9
  • Laser wavelength and sample conditioning effects on biochemical monitoring of SARS-CoV-2 VLP production upstream stage by Raman spectroscopy
    FM Dias, JP Rabello, LGO Guardalini, J Leme, TC Bernardino, L Pires, ...
    Biochemical Engineering Journal 211, 109441 , 2024
    2024
    Citations: 6
  • Monitoring by Raman spectroscopy of rabies virus‐like particles production since the initial development stages
    LGO Guardalini, RM Rangel, J Leme, TC Bernardino, SR Silveira, ...
    Journal of Chemical Technology & Biotechnology 99 (3), 658-673 , 2024
    2024
    Citations: 4
  • Sf9 cell metabolism throughout the recombinant baculovirus and rabies virus-like particles production in two Culture systems
    LGO Guardalini, J Leme, PE da Silva Cavalcante, RG de Mello, ...
    Molecular Biotechnology 66 (2), 354-364 , 2024
    2024
    Citations: 12
  • Comparison of chemometric models using Raman spectroscopy for offline biochemical monitoring throughout the VLP-making upstream process
    LGO Guardalini, VAT Dias, J Leme, TC Bernardino, RM Astray, ...
    Biochemical Engineering Journal 198, 109013 , 2023
    2023
    Citations: 19
  • Zika virus-like particles (VLPs) produced in insect cells
    RG De Mello, TC Bernardino, LGO Guardalini, RM Astray, MM Antoniazzi, ...
    Frontiers in Pharmacology 14, 1181566 , 2023
    2023
    Citations: 12
  • Sf9 cells metabolism and viability when coinfected with two monocistronic baculoviruses to produce rabies virus-like particles
    J Leme, LGO Guardalini, TC Bernardino, RM Astray, A Tonso, EGF Núñez, ...
    Molecular Biotechnology 65 (6), 970-982 , 2023
    2023
    Citations: 10

MOST CITED SCHOLAR PUBLICATIONS

  • Biochemical monitoring throughout all stages of rabies virus-like particles production by Raman spectroscopy using global models
    LGO Guardalini, PE da Silva Cavalcante, J Leme, RG de Mello, ...
    Journal of Biotechnology 363, 19-31 , 2023
    2023
    Citations: 20
  • Comparison of chemometric models using Raman spectroscopy for offline biochemical monitoring throughout the VLP-making upstream process
    LGO Guardalini, VAT Dias, J Leme, TC Bernardino, RM Astray, ...
    Biochemical Engineering Journal 198, 109013 , 2023
    2023
    Citations: 19
  • Evaluation of the applicability and effectiveness of a molecular strategy for identifying weak D and DEL phenotype among D–blood donors of mixed origin exhibiting high …
    MR Dezan, LGO Guardalini, E Pessoa, IH Ribeiro, VB Oliveira, F Luz, ...
    Transfusion 58 (2), 317-322 , 2018
    2018
    Citations: 16
  • Sf9 cell metabolism throughout the recombinant baculovirus and rabies virus-like particles production in two Culture systems
    LGO Guardalini, J Leme, PE da Silva Cavalcante, RG de Mello, ...
    Molecular Biotechnology 66 (2), 354-364 , 2024
    2024
    Citations: 12
  • Zika virus-like particles (VLPs) produced in insect cells
    RG De Mello, TC Bernardino, LGO Guardalini, RM Astray, MM Antoniazzi, ...
    Frontiers in Pharmacology 14, 1181566 , 2023
    2023
    Citations: 12
  • Performance comparison of recombinant Baculovirus and rabies virus-like particles production using two culture platforms
    LGO Guardalini, PES Cavalcante, J Leme, RG Mello, TC Bernardino, ...
    Vaccines 11 (1), 39 , 2022
    2022
    Citations: 11
  • Sf9 cells metabolism and viability when coinfected with two monocistronic baculoviruses to produce rabies virus-like particles
    J Leme, LGO Guardalini, TC Bernardino, RM Astray, A Tonso, EGF Núñez, ...
    Molecular Biotechnology 65 (6), 970-982 , 2023
    2023
    Citations: 10
  • Different modeling approaches for inline biochemical monitoring over the VLP-making upstream stages using Raman spectroscopy
    VAT Dias, LGO Guardalini, J Leme, TC Bernardino, SR da Silveira, ...
    Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 320, 124638 , 2024
    2024
    Citations: 9
  • Laser wavelength and sample conditioning effects on biochemical monitoring of SARS-CoV-2 VLP production upstream stage by Raman spectroscopy
    FM Dias, JP Rabello, LGO Guardalini, J Leme, TC Bernardino, L Pires, ...
    Biochemical Engineering Journal 211, 109441 , 2024
    2024
    Citations: 6
  • Oxygen uptake and transfer rates throughout production of recombinant baculovirus and rabies virus-like particles
    LGO Guardalini, PE da Silva Cavalcante, J Leme, RG de Mello, ...
    Process Biochemistry 124, 189-200 , 2023
    2023
    Citations: 6
  • Chemometrics and analytical blank on the at-line monitoring of Zika-VLP production using near-infrared spectroscopy
    JP Rabello, PE da Silva Cavalcante, J Leme, VAT Dias, FAC Barrence, ...
    Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 326, 125217 , 2025
    2025
    Citations: 5
  • Multiplicity of infection and culture medium on the SARS-CoV-2 virus like-particles production by baculovirus/insect system
    LG de Oliveira Guardalini, F Moura Dias, S Omae Camalhonte, J Leme, ...
    Biotechnology Letters 47 (2), 32 , 2025
    2025
    Citations: 4
  • Inline Raman spectroscopy as process analytical technology for SARS-CoV-2 VLP production
    F Moura Dias, MM Teruya, S Omae Camalhonte, V Aragão Tejo Dias, ...
    Bioprocess and Biosystems Engineering 48 (1), 63-84 , 2025
    2025
    Citations: 4
  • Monitoring by Raman spectroscopy of rabies virus‐like particles production since the initial development stages
    LGO Guardalini, RM Rangel, J Leme, TC Bernardino, SR Silveira, ...
    Journal of Chemical Technology & Biotechnology 99 (3), 658-673 , 2024
    2024
    Citations: 4
  • Biochemical monitoring of SARS-CoV-2 VLP production stages using MIR spectroscopy and chemometrics
    FM Dias, TC Bernardino, LG de Oliveira Guardalini, JL Tanaka, J Leme, ...
    Process Biochemistry 147, 418-432 , 2024
    2024
    Citations: 3
  • Raman laser intensity and sample clarification on biochemical monitoring over Zika-VLP upstream stages
    PE da Silva Cavalcante, JP Rabello, J Leme, VAT Dias, FAC Barrence, ...
    Biochemical and Biophysical Research Communications 733, 150671 , 2024
    2024
    Citations: 3
  • Scalable approach for coronavirus-like particles making based on the spike protein using Sf9 cells
    LG de Oliveira Guardalini, TC Bernardino, FM Dias, J Leme, ...
    Protein Expression and Purification 235, 106767 , 2025
    2025
    Citations: 1
  • MIR spectroscopy in ATR mode for Sf9 insect cell line biochemical monitoring in bioprocesses
    LG de Oliveira Guardalini, GM da Trindade, J Leme, TC Bernardino, ...
    Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 338, 126174 , 2025
    2025
    Citations: 1
  • Offline Raman monitoring of SARS-CoV-2 VLP upstream stage using two culture media
    J Públio Rabello, I de Sousa Fontes, F Moura Dias, S Omae Camalhonte, ...
    Spectroscopy Letters, 1-13 , 2025
    2025
    Citations: 1
  • Controlled vs. non-controlled culture systems for SARS-CoV-2 VLP production using the baculovirus/SF9 platform
    LGO Guardalini, FM Dias, TC Bernardino, J Leme, HC de Oliveira, ...
    Preparative Biochemistry & Biotechnology, 1-12 , 2026
    2026