Investigations of the Use of Beta‐propiolactone in Virus Inactivation*

LoGrippo, Gerald A.

doi:10.1111/j.1749-6632.1960.tb40931.x

Cited by 87 publications

(26 citation statements)

References 11 publications

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“…Thus, higher concentrations of BPLwere required for viral inactivation at room temperature than at 4°C. This is consistent with the increased stability of BPL at 4°C [17].…”

Section: Resultssupporting

confidence: 85%

Biosafety evaluation of recombinant protein production in goat mammary gland using adenoviral vectors: Preliminary study

Jiménez

Pose

Moltó

et al. 2012

Biotechnology Journal

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The production of recombinant proteins in the milk of non-transgenic goats can be achieved by transducing the mammary gland with recombinant adenoviral vectors. However, this process involves several regulatory issues. The current study evaluates the biosafety of this production system. We present a preliminary biosafety profile based on detection of adenoviral particles in different body fluids and the antibody response after adenoviral transduction of the goat mammary gland. In addition, two methods of adenoviral inactivation in milk were tested. Although adenoviral particles were detected in the milk until day 4 after transduction, they were absent in serum, saliva, urine and feces. Anti-adenovirus antibodies were detected in serum and milk. The virus inactivation methods neutralized adenoviral particles and preserved the immunological identity of the recombinant protein. These results support the idea of a safe production of recombinant proteins using adenoviral vectors.

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“…Thus, higher concentrations of BPLwere required for viral inactivation at room temperature than at 4°C. This is consistent with the increased stability of BPL at 4°C [17].…”

Section: Resultssupporting

confidence: 85%

Biosafety evaluation of recombinant protein production in goat mammary gland using adenoviral vectors: Preliminary study

Jiménez

Pose

Moltó

et al. 2012

Biotechnology Journal

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“…High BPL concentrations are required to reach the most buried parts of the virus, and differences in BPL diffusion within the virus may in part explain the differences reported in the literature on the BPL concentration required to inactivate a specific virus. Described as the “tailing phenomenon,” the bulk of the virus is rapidly inactivated by a small concentration of BPL, but a disproportionately larger concentration is required to inactivate the residual active virus (French, McKinney, 1964; Logrippo, 1960). Although BPL efficiency depends on the corresponding alterations of the viral structures and higher BLP concentrations will completely inactivate the virus, over-inactivation by BPL can modify viral proteins, resulting in loss or increase of antigenicity (Bonnafous et al, 2014; French, McKinney, 1964; Uittenbogaard et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

Development of an algorithm for production of inactivated arbovirus antigens in cell culture

Goodman

Russell

Velez

et al. 2014

Journal of Virological Methods

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Arboviruses are medically important pathogens that cause human disease ranging from a mild fever to encephalitis. Laboratory diagnosis is essential to differentiate arbovirus infections from other pathogens with similar clinical manifestations. The Arboviral Diseases Branch (ADB) reference laboratory at the CDC Division of Vector-Borne Diseases (DVBD) produces reference antigens used in serological assays such as the virus-specific immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). Antigen production in cell culture has largely replaced the use of suckling mice; however, the methods are not directly transferable. The development of a cell culture antigen production algorithm for nine arboviruses from the three main arbovirus families, Flaviviridae, Togaviridae, and Bunyaviridae, is described here. Virus cell culture growth and harvest conditions were optimized, inactivation methods were evaluated, and concentration procedures were compared for each virus. Antigen performance was evaluated by the MAC-ELISA at each step of the procedure. The antigen production algorithm is a framework for standardization of methodology and quality control; however, a single antigen production protocol was not applicable to all arboviruses and needed to be optimized for each virus.

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“…The combination of methylene blue and white light irradiation effectively inactivated the Ad-E2 vector initially present in the first samples of milk collected. With this approach, the use of inactivating agents such as the potentially toxic beta-propiolactone was avoided, in contrast with other procedures previously proposed (25,26).…”

Section: Discussionmentioning

confidence: 99%

Effectiveness of the E2-Classical Swine Fever Virus Recombinant Vaccine Produced and Formulated within Whey from Genetically Transformed Goats

Sánchez¹,

Barrera²,

Farnós³

et al. 2014

Clin. Vaccine Immunol.

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e Subunit recombinant vaccines against classical swine fever virus (CSFV) are a promising alternative to overcome practical and biosafety issues with inactivated vaccines. One of the strategies in evaluation under field conditions is the use of a new marker E2-based vaccine produced in the milk of adenovirally transduced goats. Previously we had demonstrated the efficacy of this antigen, which conferred early protection and long-lasting immunity in swine against CSFV infection. Here, we have used a simpler downstream process to obtain and formulate the recombinant E2 glycoprotein expressed in the mammary gland. The expression levels reached approximately 1.7 mg/ml, and instead of chromatographic separation of the antigen, we utilized a clarification process that eliminates the fat content, retains a minor amount of caseins, and includes an adenoviral inactivation step that improves the biosafety of the final formulation. In a vaccination and challenge experiment in swine, different doses of the E2 antigen contained within the clarified whey generated an effective immune response of neutralizing antibodies that protected all of the animals against a lethal challenge with CSFV. During the immunization and after challenge, the swine were monitored for adverse reactions related to the vaccine or symptoms of CSF, respectively. No adverse reactions or clinical signs of disease were observed in vaccinated animals, in which no replication of CSFV could be detected after challenge. Overall, we consider that the simplicity of the procedures proposed here is a further step toward the introduction and implementation of a commercial subunit vaccine against CSF.

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Investigations of the Use of Beta‐propiolactone in Virus Inactivation*

Cited by 87 publications

References 11 publications

Biosafety evaluation of recombinant protein production in goat mammary gland using adenoviral vectors: Preliminary study

Biosafety evaluation of recombinant protein production in goat mammary gland using adenoviral vectors: Preliminary study

Development of an algorithm for production of inactivated arbovirus antigens in cell culture

Effectiveness of the E2-Classical Swine Fever Virus Recombinant Vaccine Produced and Formulated within Whey from Genetically Transformed Goats

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