Preclinical optimization of an enterotoxigenic Escherichia coli adjuvanted subunit vaccine using response surface design of experiments

Poncet, David; Hessler, Catherine; Liang, Hong; Gautheron, Sylviane; Sergent, Michelle; Rintala, Nicholas D.; Seydoux, Emilie; Huang, Po‐Wei; Argilla, David; Ruiz, Sophie; Heinrichs, Jon H.; Maciel, Milton; Orr, Mark T.

doi:10.1038/s41541-020-00228-w

Cited by 9 publications

(8 citation statements)

References 55 publications

(71 reference statements)

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“…According to the quality by design (QbD) framework, to obtain a robust adenovirus production process, a robust setpoint and design space for the process parameters should be explored using the design of experiment (DoE) approach (Zhang and Mao 2017 ). This strategy is available for antibodies and virus vaccines (Patel et al 2018 ; Poncet et al 2020 ) but has seldom been reported for adenovirus production.…”

Section: Introductionmentioning

confidence: 99%

Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production

et al. 2022

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Herpes zoster is caused by reactivation of the varicella zoster virus (VZV). Researching and developing a herpes zoster vaccine will help to decrease the incidence of herpes zoster. To increase the bioreactor productivity, a serum-free HEK293 cell perfusion process with adenovirus vector herpes zoster (rAd-HZ) vaccine production was developed efficiently using the design of experiment (DoE) method. First, serum-free media for HEK293 cells were screened in both batch and semi-perfusion culture modes. Then, three optimal media were employed in a medium mixture design to improve cell culture performance, and the 1:1 mixture of HEK293 medium and MCD293 medium (named HM293 medium) was identified as the optimal formulation. On the basis of the HM293 medium, the relationship of critical process parameters (CPPs), including the time of infection (TOI), multiplicity of infection (MOI), pH, and critical quality attributes (CQAs) (adenovirus titer (Titer), cell-specific virus yield (CSVY), adenovirus fold expansion (Fold)) of rAd-HZ production was investigated using the DoE approach. Furthermore, the robust setpoint and design space of these CPPs were explored. Finally, the rAd-HZ production process with parameters at a robust setpoint (TOI = 7.2 × 106 cells/mL, MOI = 3.7, and pH = 7.17) was successfully scaled-up to a 3-L bioreactor with an alternating tangential flow system, yielding an adenovirus titer of 3.0 × 1010 IFU/mL, a CSVY of 4167 IFU/cells, a Fold of 1117 at 2 days post infection (dpi). The DoE approach accelerated the development of a HEK293 serum-free medium and of a robust adenovirus production process.

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Section: Introductionmentioning

confidence: 99%

Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production

et al. 2022

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“…Two separate immunogenicity experiments were conducted based on DOE and/or desirability index methodology adapted from a previous report (3). The first immunogenicity experiment focused on optimization of vaccine formulation component doses (LecA antigen, GLA, 3M-052, phospholipid) using a central composite DOE.…”

Section: Resultsmentioning

confidence: 99%

“…For instance, the association of stool IgA and PBMC IFNg with protection from amebiasis in humans informed the weighting factors of the immunological readouts employed in the DOE and desirability index analysis here. Even when correlates of immunity are known, the weighting of the various readouts will always have some degree of subjectivity and may require refinement as more knowledge is gained (3). Another potential benefit of the DOE and desirability index approach is identification of candidates that elicit different immune response profiles.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, immunology assays enable a wide range of parameters to be monitored, each of which may be more or less dependent on the individual vaccine components. Despite this complexity, efficient dose and formulation optimization approaches employing tools such as Design of Experiments (DOE) or desirability functions are only rarely reported in vaccine development (1)(2)(3). Nevertheless, such approaches offer the benefit of objective multifactorial analysis while reducing the number of experimental subjects, such as animals, and the number of tests that would otherwise be required.…”

Section: Introductionmentioning

confidence: 99%

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Optimizing a Multi-Component Intranasal Entamoeba Histolytica Vaccine Formulation Using a Design of Experiments Strategy

et al. 2021

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Amebiasis is a neglected tropical disease caused by Entamoeba histolytica. Although the disease burden varies geographically, amebiasis is estimated to account for some 55,000 deaths and millions of infections globally per year. Children and travelers are among the groups with the greatest risk of infection. There are currently no licensed vaccines for prevention of amebiasis, although key immune correlates for protection have been proposed from observational studies in humans. We previously described the development of a liposomal adjuvant formulation containing two synthetic TLR ligands (GLA and 3M-052) that enhanced antigen-specific fecal IgA, serum IgG2a, a mixed IFNγ and IL-17A cytokine profile from splenocytes, and protective efficacy following intranasal administration with the LecA antigen. By applying a statistical design of experiments (DOE) and desirability function approach, we now describe the optimization of the dose of each vaccine formulation component (LecA, GLA, 3M-052, and liposome) as well as the excipient composition (acyl chain length and saturation; PEGylated lipid:phospholipid ratio; and presence of antioxidant, tonicity, or viscosity agents) to maximize desired immunogenicity characteristics while maintaining physicochemical stability. This DOE/desirability index approach led to the identification of a lead candidate composition that demonstrated immune response durability and protective efficacy in the mouse model, as well as an assessment of the impact of each active vaccine formulation component on protection. Thus, we demonstrate that both GLA and 3M-052 are required for statistically significant protective efficacy. We also show that immunogenicity and efficacy results differ in female vs male mice, and the differences appear to be at least partly associated with adjuvant formulation composition.

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“…This tip adhesin complex was subsequently tested for immunogenicity when delivered transcutaneously with mutant LT (R192G) via patch in human volunteers (87). Although these studies indicate that additional effort will be required for optimal delivery of the fimbrial tip adhesin complex (121), recent studies of dscCfaE genetically fused in a chimeric complex with the A2 subunit of cholera toxin and the B subunit of LT (dscCfaE-sCTA2/LTB) resulted in substantial protection in an Aotus nancymaae model of infection, following intradermal vaccination with mLT(R192G) (89). Similar preclinical studies of CS6, an antigen present in~1/5 of ETEC strains have been ongoing.…”

Section: Development Of Cf and Cs Subunit Vaccinesmentioning

confidence: 99%

Confronting Challenges to Enterotoxigenic Escherichia coli Vaccine Development

Fleckenstein

2021

Front. Trop. Dis

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The enterotoxigenic Escherichia coli (ETEC) are a diverse and genetically plastic pathologic variant (pathovar) of E. coli defined by their production of heat-labile (LT) and heat-stable (ST) enterotoxins. These pathogens, which came to recognition more than four decades ago in patients presenting with severe cholera-like diarrhea, are now known to cause hundreds of millions of cases of symptomatic infection annually. Children in low-middle income regions of the world lacking access to clean water and basic sanitation are disproportionately affected by ETEC. In addition to acute diarrheal morbidity, these pathogens remain a significant cause of mortality in children under the age of five years and have also been linked repeatedly to sequelae of childhood malnutrition and growth stunting. Vaccines that could prevent ETEC infections therefore remain a high priority. Despite several decades of effort, a licensed vaccine that protects against the breadth of these pathogens remains an aspirational goal, and the underlying genetic plasticity of E. coli has posed a fundamental challenge to development of a vaccine that can encompass the complete antigenic spectrum of ETEC. Nevertheless, novel strategies that include toxoids, a more complete understanding of ETEC molecular pathogenesis, structural details of target immunogens, and the discovery of more highly conserved antigens essential for virulence should accelerate progress and make a broadly protective vaccine feasible.

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Preclinical optimization of an enterotoxigenic Escherichia coli adjuvanted subunit vaccine using response surface design of experiments

Cited by 9 publications

References 55 publications

Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production

Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production

Optimizing a Multi-Component Intranasal Entamoeba Histolytica Vaccine Formulation Using a Design of Experiments Strategy

Confronting Challenges to Enterotoxigenic Escherichia coli Vaccine Development

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