Virus-like particle bioprocessing: challenges and opportunities

Middelberg, Anton P. J.; Lua, Linda H.L.

doi:10.4155/pbp.13.49

Cited by 5 publications

(5 citation statements)

References 20 publications

(23 reference statements)

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“…We found that L1:P18I10 VLPs formed a distinctive band at the interface layer between 20% and 70% sucrose. This pattern was well in line with previous studies observing that HPV16 L1 VLPs form a visible band at a concentration of 30-40% in a continuous sucrose gradient [41]. As the HPV16 L1 VLPs are hollow interior and might have DNA-capsid affinity [78,79], SC-purified L1:P18I10 VLPs might encapsulate DNA and lead to irregular or heterogeneous forms [80].…”

Section: Discussionsupporting

confidence: 92%

“…In our laboratory, the Sf9 cell density at 96 h of harvest could only reach approximately 1.2 to 1.5 × 10 6 cells/mL. This pattern match Merck's application note indicating that the Sf9 cell density could expand from 1.0-1.2 × 10 6 cells/mL to 1.5-2 × 10 6 cells/mL through shake flask cultures to a bioreactor [41]. However, some studies reported that the density of Sf9 cells could reach over 10 × 10 6 cells/mL by using fed-batch bioreactors under tightly monitored culture conditions for large-scale manufacturing production [37].…”

Section: Discussionsupporting

confidence: 76%

“…Therefore, purified L1:P18I10 VLPs should undergo baculovirus inactivation to eradicate the potential infectivity [71] or baculovirus removal through extra chromatographic steps. For instance, the ion exchange chromatography was shown to remove 10 2 to 10 5 baculovirus particles during VLP purification [41].…”

Section: Discussionmentioning

confidence: 99%

“…The BEVS/IC system has advantages over the mammalian expression system to reach a high expression level of recombinant proteins that is comparable with bacteria and yeast expression systems [40]. The disadvantage of the BEVS/IC platform could attribute to the baculoviruses that must be inactivated or removed through extra downstream steps, such as chromatography [41]. Although the mammalian cell expression system provides a baculovirus-free purification condition, low expression level and high cost could be its major drawbacks [36].…”

Section: Introductionmentioning

confidence: 99%

“…However, the conventional ultracentrifugation procedures were quite time-consuming and difficult for industrial scaling-up. A large proportion of target protein was lost during purification, and the recovery rate (~10%) was relatively low [41]. A substantial quantity and quality of VLP-based vaccine is necessary for biophysical characterization and downstream immunogenic test.…”

Section: Introductionmentioning

confidence: 99%

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Chimeric Human Papillomavirus-16 Virus-like Particles Presenting HIV-1 P18I10 Peptide: Expression, Purification, Bio-Physical Properties and Immunogenicity in BALB/c Mice

Chen

Saubí

Joseph-Munné

2023

IJMS

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Human papillomavirus (HPV) vaccines based on HPV L1 virus-like particles (VLPs) are already licensed but not accessible worldwide. About 38.0 million people were living with HIV in 2020 and there is no HIV vaccine yet. Therefore, safe, effective, and affordable vaccines against both viruses are an urgent need. In this study, the HIV-1 P18I10 CTL peptide from the V3 loop of HIV-1 gp120 glycoprotein was inserted into the HPV16 L1 protein to construct chimeric HPV:HIV (L1:P18I10) VLPs. Instead of the traditional baculovirus expression vector/insect cell (BEVS/IC) system, we established an alternative mammalian 293F cell-based expression system using cost-effective polyethylenimine-mediated transfection for L1:P18I10 protein production. Compared with conventional ultracentrifugation, we optimized a novel chromatographic purification method which could significantly increase L1:P18I10 VLP recovery (~56%). Chimeric L1:P18I10 VLPs purified from both methods were capable of self-assembling to integral particles and shared similar biophysical and morphological properties. After BALB/c mice immunization with 293F cell-derived and chromatography-purified L1:P18I10 VLPs, almost the same titer of anti-L1 IgG (p = 0.6409) was observed as Gardasil anti-HPV vaccine-immunized mice. Significant titers of anti-P18I10 binding antibodies (p < 0.01%) and P18I10-specific IFN-γ secreting splenocytes (p = 0.0002) were detected in L1:P18I10 VLP-immunized mice in comparison with licensed Gardasil-9 HPV vaccine. Furthermore, we demonstrated that insertion of HIV-1 P18I10 peptide into HPV16 L1 capsid protein did not affect the induction in anti-L1 antibodies. All in all, we expected that the mammalian cell expression system and chromatographic purification methods could be time-saving, cost-effective, scalable platforms to engineer bivalent VLP-based vaccines against HPV and HIV-1.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Chimeric Human Papillomavirus-16 Virus-like Particles Presenting HIV-1 P18I10 Peptide: Expression, Purification, Bio-Physical Properties and Immunogenicity in BALB/c Mice

Chen

Saubí

Joseph-Munné

2023

IJMS

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Current status and future challenges in transitioning to continuous bioprocessing of virus‐like particles

Mittal

Banerjee

Lua

et al. 2021

J of Chemical Tech & Biotech

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Virus-like particles (VLPs) are bioengineered protein complexes that can serve as prophylactic vaccines due to favorable immunological characteristics like size, repetitive surface geometry and ability to induce both innate and adaptive immune responses, without being infectious. Large-scale manufacturing of VLPs presents some interesting and unique challenges associated with removal of refractory contaminants to attain high product purity, improvement in structural homogeneity and colloidal stability. Recent advancements in production and purification of VLP-based vaccines, such as application of monolithic multicolumn chromatography and inline concentration and diafiltration, have set a stage for implementation of continuous bioprocessing for commercial manufacturing of VLPs. The benefits of continuous bioprocessing have been well demonstrated for biotherapeutics, particularly monoclonal antibodies and small peptides. With the growing demand for affordable vaccines, the biopharmaceutical industry has shown increasing interest in establishing end-to-end continuous platforms. This article reviews major developments that have occurred in continuous processing for the manufacturing of VLPs. The major problems inherent in transitions from batch to continuous processing are also discussed, along with potential strategies to overcome the challenges.

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Purification of flavivirus VLPs by a two-step chomatographic process

Lima

Souza

Castilho

2019

Vaccine

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Virus-like particle bioprocessing: challenges and opportunities

Abstract: Rapid advances in the design and manufacture of structured materials will continue to see high levels of innovation, and the commercialization of new matrices tailored to virus-like particle purification."

Cited by 5 publications

References 20 publications

Chimeric Human Papillomavirus-16 Virus-like Particles Presenting HIV-1 P18I10 Peptide: Expression, Purification, Bio-Physical Properties and Immunogenicity in BALB/c Mice

Chimeric Human Papillomavirus-16 Virus-like Particles Presenting HIV-1 P18I10 Peptide: Expression, Purification, Bio-Physical Properties and Immunogenicity in BALB/c Mice

Current status and future challenges in transitioning to continuous bioprocessing of virus‐like particles

Purification of flavivirus VLPs by a two-step chomatographic process

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