Inactivation of mycoplasmas by use of phenol, formalin and beta-propiolactone

Koski, T. A.; Christianson, Gwenn; Cole, F.L.

doi:10.1016/0092-1157(76)90026-3

Cited by 7 publications

(4 citation statements)

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“…The results of our study using formaldehyde and BPL were consistent with previous data obtained for these reagents using only three mycoplasma species (M. gallisepticum, M. canis, and A. laidlawii) spiked into Vero and DK cell culture suspensions (30). Similar to the results described in the previous study, we observed incomplete inactivation of many of the species tested when the concentration of formaldehyde was 0.02% or lower.…”

Section: Discussionsupporting

confidence: 82%

“…A. laidlawii was also found to be more resistant to BPL than the other species examined. A. laidlawii remained viable for 2 h of incubation with 0.1% BPL; this result is consistent with the ability of this species to resist inactivation by BPL described previously (30). In a majority of the protocols used for production of inactivated virus vaccines (2,9,17,25,27,29,33,42,51,53), the concentration of BPL and formaldehyde exceeds the critical concentration (0.1% to 0.2%) required for complete inactivation of mycoplasmas.…”

Section: Discussionsupporting

confidence: 69%

“…Similar to the results described in the previous study, we observed incomplete inactivation of many of the species tested when the concentration of formaldehyde was 0.02% or lower. A significant improvement in inactivation of mycoplasmas was observed when the formaldehyde concentration was increased (30). Our data also showed that A. laidlawii exhibited enhanced stability in the presence of formaldehyde (30) and resistance to inactivation for the first 5 h of incubation with 0.2% formaldehyde.…”

Section: Discussionsupporting

confidence: 62%

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Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

David

Volokhov

et al. 2010

Appl Environ Microbiol

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Although mycoplasmas are generally considered to be harmless commensals, some mycoplasma species are able to cause infections in pediatric, geriatric, or immunocompromised patients. Thus, accidental contamination of biologics with mycoplasmas represents a potential risk for the health of individuals who receive cell-derived biological and pharmaceutical products. To assess the efficiency of inactivation of mycoplasmas by the agents used in the manufacture of egg-derived influenza vaccines, we carried out a series of experiments aimed at monitoring the viability of mycoplasmas spiked into both chicken allantoic fluid and protein-rich microbiological media and then treated with beta-propiolactone, formalin, cetyltrimethylammonium bromide, Triton X-100, and sodium deoxycholate, which are agents that are commonly used for virus inactivation and disruption of viral particles during influenza vaccine production. Twenty-two mycoplasma species (with one to four strains of each species) were exposed to these inactivating agents at different concentrations. The most efficient inactivation of the mycoplasmas evaluated was observed with either 0.5% Triton X-100 or 0.5% sodium deoxycholate. Cetyltrimethylammonium bromide at concentrations of >0.08% was also able to rapidly inactivate (in less than 30 min) all mycoplasmas tested. In contrast, negligible reductions in mycoplasma titers were observed with 0.0125 to 0.025% formaldehyde. However, increasing the concentration of formaldehyde to 0.1 to 0.2% improved the mycoplasmacidal effect. Incubation of mycoplasmas with 0.1% beta-propiolactone for 1 to 24 h had a marked mycoplasmacidal effect. A comparison of the mycoplasma inactivation profiles showed that strains of selected species (Mycoplasma synoviae, Mycoplasma gallisepticum, Mycoplasma orale, Mycoplasma pneumoniae, and Acholeplasma laidlawii) represent a set of strains that can be utilized to validate the effectiveness of mycoplasma clearance obtained by inactivation and viral purification processes used for the manufacture of an inactivated egg-based vaccine.The mycoplasmas (the trivial name for Mollicutes) are a class of broadly distributed wall-less bacteria with some of the smallest known genomes that support bacterial self-replication (43). While most mycoplasmas are naturally harmless commensals that colonize a wide range of plant, insect, reptilian, avian, mammalian, and human hosts (43), some of them are able to cause diseases whose severity varies in their natural hosts (1,4,10,19,43).Mycoplasmas, particularly species of genera Mycoplasma and Acholeplasma, are also frequent contaminants of primary and continuous cell lines (45). This is a concern for research laboratories and commercial facilities involved in the development and production of cell-derived biological and pharmaceutical products. In addition, several natural mycoplasma infections of embryonated fowl eggs and primary chicken embryo cell cultures used for the production of viral vaccines have been reported (5-7, 16, 36, 46, 48, 55). Although only a ...

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Section: Discussionsupporting

confidence: 82%

Section: Discussionsupporting

confidence: 69%

Section: Discussionsupporting

confidence: 62%

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Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

David

Volokhov

et al. 2010

Appl Environ Microbiol

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“…After inactivation process the bacteria were washed with sterile buffer saline water to remove protein contamination. To ascertain the completion of inactivation process, the inactivated broth was separately cultured on Mycoplasma broth and agar, incubated at 37˚C at 10% CO 2 tension and observed for seven days for appearance of any specific growth of Mycoplasma colonies or color change, respectively [17]. After inactivating the final product, there was no evidence of growth of Mycoplasma observed.…”

Section: Inactivation Of Killed Vaccinementioning

confidence: 99%

Inactivated Vaccine Trial of <i>Mycoplasma gallisepticum</i> in Ethiopia

Bekele¹,

Assefa²

2018

OJVM

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Derivatization of β-propiolactone in biological mixtures for enhanced gas chromatographic mass spectrometric determinations

Phillips

Fraser

1981

Biol. Mass Spectrom.

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Detection and identification of low levels (<200 ppb) of free P-propiolactone in complex biological mixtures were achieved through in siru chemical derivatization and subsequent gas chromatographic mass spectrometric determination of the products. P-Propiolactone, when reacted with octadecylamine, followed by trimethylsilylation, yielded a derivative which exhibited both good gas chromatographic characteristics and highly interpretable mass spectrometric data.

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Inactivation of mycoplasmas by use of phenol, formalin and beta-propiolactone

Cited by 7 publications

References 2 publications

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

Inactivated Vaccine Trial of <i>Mycoplasma gallisepticum</i> in Ethiopia

Derivatization of β-propiolactone in biological mixtures for enhanced gas chromatographic mass spectrometric determinations

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Inactivation of mycoplasmas by use of phenol, formalin and beta-propiolactone

Cited by 7 publications

References 2 publications

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model

Inactivated Vaccine Trial of &lt;i&gt;Mycoplasma gallisepticum&lt;/i&gt; in Ethiopia

Derivatization of β-propiolactone in biological mixtures for enhanced gas chromatographic mass spectrometric determinations

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Inactivated Vaccine Trial of <i>Mycoplasma gallisepticum</i> in Ethiopia