Novel constructs and 1-step chromatography protocols for the production of Porcine Circovirus 2d (PCV2d) and Circovirus 3 (PCV3) subunit vaccine candidates

Peswani, Amber R; Narkpuk, Jaraspim; Krueger, A. P.; Bracewell, Daniel G.; Lekcharoensuk, Porntippa; Haslam, Stuart M.; Dell, Anne; Jaru-Ampornpan, Peera; Robinson, Colin

doi:10.1016/j.fbp.2021.10.001

Cited by 9 publications

(15 citation statements)

References 26 publications

(34 reference statements)

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“…As we observed impurities at 200 mM NaCl, we maintained the equilibration buffer at 300 mM NaCl. This optimization resulted in improved binding of PCV2d (by 17%) compared to the previously reported process [ 22 ]. Therefore, these conditions were applied to the HiScale 16/40 experiments, where the linear velocity was kept constant at 300 cm/h.…”

Section: Resultsmentioning

confidence: 95%

“…The VN titers for pre-immunization sera were insignificant. This convinced them that the construct shows high potential for being further developed as a subunit vaccine against PCV2d, and hence that proceeding to large-scale process development should be carried-out [ 22 ].…”

Section: Resultsmentioning

confidence: 99%

“…Cell stocks of E. coli W3110 containing pET23/ptac Δ2–40 PCV2d-His6 were received from the Robinson group at the University of Kent (UK) [ 22 ]. Research cell banks (RCB) were prepared by growing cells to an OD600 of 0.5–0.7 in Terrific Broth (TB) (BD Difco TM , Sparks, NV, USA) containing 5 g/L glycerol and 100 µg/mL ampicillin at 30 °C and 200 rpm in a shaking incubator (Innova43R, Eppendorf New Brunswick, Enfield, CT, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, a novel PCV2d vaccine and a laboratory-scale production process were developed in order to produce small quantities of test material for animal experiments and vaccine characterization [ 22 ]. However, for pig trials and manufacturing purposes, the development of a larger-scale production process is needed.…”

Section: Introductionmentioning

confidence: 99%

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A Simple and Cost-Efficient Platform for a Novel Porcine Circovirus Type 2d (PCV2d) Vaccine Manufacturing

et al. 2023

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Porcine circovirus type 2d (PCV2d) is becoming the predominant PCV genotype and considerably affects the global pig industry. Nevertheless, currently, no commercial PCV2d vaccine is available. Preventing and controlling the disease caused by PCV2d is therefore based on other genotype-based vaccines. However, their production platforms are laborious, limited in expression level, and relatively expensive for veterinary applications. To address these challenges, we have developed a simple and cost-efficient platform for a novel PCV2d vaccine production process, using fed-batch E. coli fermentation followed by cell disruption and filtration, and a single purification step via cation exchange chromatography. The process was developed at bench scale and then pilot scale, where the PCV2d subunit protein yield was approximately 0.93 g/L fermentation volume in a short production time. Moreover, we have successfully implemented this production process at two different sites, in Southeast Asia and Europe. This demonstrates transferability and the high potential for successful industrial production.

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Section: Resultsmentioning

confidence: 95%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Simple and Cost-Efficient Platform for a Novel Porcine Circovirus Type 2d (PCV2d) Vaccine Manufacturing

et al. 2023

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“…However, low levels of PCV2 viremia mean that the virus has not completely cleared, resulting in chronic subclinical infection with PCV2. Therefore, these commercial PCV2 vaccines appear to provide incomplete cross-protection against the current dominant PCV2d genotype ( Peswani et al, 2022 ). Consequently, it is important that next-generation PCV2 vaccines induce sufficient immune protection against PCV2d.…”

Section: Discussionmentioning

confidence: 99%

A porcine circovirus type 2d-based virus-like particle vaccine induces humoral and cellular immune responses and effectively protects pigs against PCV2d challenge

Kim,

Choi,

Shin

et al. 2024

Front. Microbiol.

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The pathogenic porcine circovirus type 2 (PCV2) leads to significant economic losses in pig production. PCV2d is currently the dominant genotype causing porcine circovirus-associated disease (PCVAD) worldwide. Therefore, development of a recombinant PCV2d-based vaccine is required to elicit complete protection against PCV2d infection. In this study, we generated virus-like particles of PCV2d-based capsid protein (Bac-2dCP) using a baculovirus expression system and evaluated its protective efficacy against PCV2d infection in specific pathogen-free (SPF) pigs. Three-week-old SPF miniature pigs were intramuscularly immunized with purified Bac-2dCP and intranasally challenged with PCV2d at 4 weeks post-vaccination. The Bac-2dCP group showed significantly higher IgG levels and neutralizing antibodies against PCV2b and PCV2d genotypes, as well as increased interferon-γ levels, and increased body weight and average daily weight gain compared with positive (challenged) and negative (unchallenged) controls. In particular, the Bac-2dCP group showed almost complete absence of PCV2d DNA in serum, nasal, and rectal swabs and in lung, lymph node, and kidney tissue samples. However, the positive control group exhibited low levels of neutralizing antibody, and high levels of PCV2 DNA in serum, swab, and tissue samples, resulting in PCV2-associated pathological lesions. The results of this study demonstrated that a recombinant Bac-2dCP vaccine conferred complete protection against a PCV2d challenge in SPF miniature pigs.

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Coinfections of African swine fever virus, porcine circovirus 2 and 3, and porcine parvovirus 1 in swine in Nigeria

et al. 2022

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Novel constructs and 1-step chromatography protocols for the production of Porcine Circovirus 2d (PCV2d) and Circovirus 3 (PCV3) subunit vaccine candidates

Cited by 9 publications

References 26 publications

A Simple and Cost-Efficient Platform for a Novel Porcine Circovirus Type 2d (PCV2d) Vaccine Manufacturing

A Simple and Cost-Efficient Platform for a Novel Porcine Circovirus Type 2d (PCV2d) Vaccine Manufacturing

A porcine circovirus type 2d-based virus-like particle vaccine induces humoral and cellular immune responses and effectively protects pigs against PCV2d challenge

Coinfections of African swine fever virus, porcine circovirus 2 and 3, and porcine parvovirus 1 in swine in Nigeria

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