Development of a Drosophila S2 insect-cell based placental malaria vaccine production process

Jongh, Wian A de; Resende, Mafalda; Leisted, Carsten; Strøbæk, Anette; Berisha, Besim; Nielsen, Morten A; Salanti, Ali; Hjerrild, Kathryn A.; Draper, Simon J.; Dyring, Charlotte

doi:10.1186/1753-6561-7-s6-p20

Cited by 4 publications

(4 citation statements)

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“…As a result of these advantages, S2 cells are being used for a variety of purposes, including the production of vaccine antigens and other heterologous proteins. It is noteworthy that the S2 system may allow for higher yields of these proteins than bacterial and yeast expression systems ( 28, 29 ).…”

Section: Introductionmentioning

confidence: 99%

Expression and purification of HER2 extracellular domain proteins in Schneider2 insect cells

Kanthala

Mill

Riese

et al. 2016

Protein Expression and Purification

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Overexpression of human epidermal growth factor receptor 2 (HER2/ErbB2/Neu) results in ligand independent activation of kinase signaling and is found in about 30% of human breast cancers, and is correlated with a more aggressive tumor phenotype. The HER2 extracellular domain (ECD) consists of four domains − I, II, III and IV. Although the role of each domain in the dimerization and activation of the receptor has been extensively studied, the role of domain IV (DIV) is not clearly understood yet. In our previous studies, we reported peptidomimetic molecules inhibit HER2:HER3 heterodimerization. In order to study the binding interactions of peptidomimetics with HER2 DIV in detail, properly folded recombinant HER2 protein in pure form is important. We have expressed and purified HER2 ECD and DIV proteins in the Drosophila melanogaster Schneider2 (S2) cell line. Using the commercial Drosophila expression system (DES), we transfected S2 cells with plasmids designed to direct the expression of secreted recombinant HER2 ECD and DIV proteins. The secreted proteins were purified from the conditioned medium by filtration, ultrafiltration, dialysis and nickel affinity chromatography techniques. The purified HER2 proteins were then analyzed using western blot, mass spectrometry and circular dichroism (CD) spectroscopy.

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Section: Introductionmentioning

confidence: 99%

Expression and purification of HER2 extracellular domain proteins in Schneider2 insect cells

Kanthala

Mill

Riese

et al. 2016

Protein Expression and Purification

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“…The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) was expressed in S2 cells of Drosophila to produce non-glycosylated variants capable of binding to its receptor in rabbits resulted in IgG production against PfRH5 protein [112]. Highly potential vaccine VAR2CSA against malaria was successfully produced in S2 cells of Drosophila melanogaster [113].…”

Section: In-vitro Model Of Epidemic and Pandemic Infectious Diseasementioning

confidence: 99%

Drosophila melanogaster: A Robust Tool to Study Candidate Drug against Epidemic and Pandemic Diseases

Samadder¹

2020

Animal Models in Medicine and Biology

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Drosophila melanogaster is a widely used, dynamic model organism to study various pathogenic diseases observed ubiquitously in the human population. Drosophila, at present, is extensively used to conduct preclinical studies besides its counterpart rodents. The epidemic and pandemic diseases are discussed in this review to demonstrate Drosophila melanogaster as a key model. Epidemic and pandemic diseases are still claiming more than 5 million lives every year, and these diseases were well studied in flies. Currently there is no cure for the disease like HIV; the bacterial and fungal infections usually seen in HIV/AIDS patients could be demonstrated elaborately in Drosophila melanogaster. Diseases like myocardial infractions and cancer causing viral infection are long term effects of ART (antiretroviral therapy) that could be experimented in flies. Stable Drosophila S2 cell line, Transgenic flies, transfusion of bacteria and fungi could be implemented to study several infectious diseases and for vaccine development. The latest trends in understanding pathogenic diseases and its potential biochemical markers in flies are discussed in this review to utilize the fruit flies as a functional tool and to explore further it in drug development. The advantages and disadvantages of the fly as a model of infection are discussed along with the epidemiology and the cellular pathophysiology

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“…Various processes for bulk manufacturing of viral-based vaccines were presented, for example, three strains of H1N1, H3N2, and influenza B vaccines were cultured in Cytodex microcarriers, fed-batch culture, and perfusion bioreactor equipped with a spin filter, respectively, , and influenza A virus was expanded in the perfusion bioreactor using an alternating tangential flow (ATF) . Also, disposable fixed-bed bioreactors have been used in the adherent cell culture technique to produce a higher yield of live viruses , and there are various studies showing a comparable cell growth rate and virus production yield for disposable and stainless-steel bioreactors . Cell culture-based vaccine production employing bioreactors has expanded manufacturing capacities and facilitated improved process control, while ensuring the product quality.…”

Section: Introductionmentioning

confidence: 99%

“… 25 Also, disposable fixed-bed bioreactors have been used in the adherent cell culture technique to produce a higher yield of live viruses 26 , 27 and there are various studies showing a comparable cell growth rate and virus production yield for disposable and stainless-steel bioreactors. 28 Cell culture-based vaccine production employing bioreactors has expanded manufacturing capacities and facilitated improved process control, while ensuring the product quality. Technology based on cell culture gives producers the ability to adapt to market requirements more rapidly with shorter production cycles.…”

Section: Introductionmentioning

confidence: 99%

High-Titer Recombinant Adenovirus 26 Vector GMP Manufacturing in HEK 293 Cells with a Stirred Single-Use Bioreactor for COVID-19 Vaccination Purposes

Sedighikamal,

Sattarzadeh,

Karimi Mostofi

et al. 2023

ACS Omega

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The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 virus) pandemic has shown the importance of pursuing various vaccine manufacturing strategies. In the present study, the HEK 293 cells were infected with recombinant adenovirus serotype 26 (rAd26), and the effects of critical process parameters (CPPs) including viable cell density (VCD) at infection time (0.5 × 10 6 , 0.8 × 10 6 , 1.4 × 10 6 , 1.8 × 10 6 , and 2.5 × 10 6 cells/mL), the multiplicity of infection (MOI) = 3, 6, 9, 12, and 15, and two aeration strategies (high-speed agitation with a sparging system and low-speed agitation with an overlay system) were investigated experimentally. The results of small-scale experiments in 2 L shake flasks (SF 2L) demonstrated that the initial VCD and MOI could affect the cell proliferation and viability. The results at these experiments showed that VCD = 1.4 × 10 6 cells/mL and MOI = 9 yielded TCID 50 /mL = 10 8.9 , at 72 h post-infection (hpi), while the virus titer at VCD = 0.5 × 10 6 and 0.8 × 10 6 cells/mL was lower compared to that of VCD = 1.4 × 10 6 cells/mL. Moreover, our findings showed that VCDs > 1.8 × 10 6 cells/m with MOI = 9 did not have a positive effect on TCID 50 /mL and MOI = 3 and 6 were less efficient, whereas MOI > 12 decreased the viability drastically. In the next step, the optimized CPPs in a small scale were exploited in a 200 L single-use bioreactor (SUB), with good manufacturing practice (GMP) conditions, at RPM = 25 with an overlay system, yielding high-titer rAd26 manufacturing, i.e., TCID 50 /mL = 10 8.9 , at 72 hpi.

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Development of a Drosophila S2 insect-cell based placental malaria vaccine production process

Cited by 4 publications

References 0 publications

Expression and purification of HER2 extracellular domain proteins in Schneider2 insect cells

Expression and purification of HER2 extracellular domain proteins in Schneider2 insect cells

Drosophila melanogaster: A Robust Tool to Study Candidate Drug against Epidemic and Pandemic Diseases

High-Titer Recombinant Adenovirus 26 Vector GMP Manufacturing in HEK 293 Cells with a Stirred Single-Use Bioreactor for COVID-19 Vaccination Purposes

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