Scientific Publications by FDA Staff
Appl Environ Microbiol 2010 May;76(9):2718-28
Evaluation of Mycoplasma inactivation during production of biologics: egg-based viral vaccines as a model.
David SA, Volokhov DV, Zhiping Y, Chizhikov V
Although mycoplasmas are generally considered to be harmless commensals, some mycoplasma species are able to cause infections among 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 mycoplasma inactivation 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 followed by treatment with beta-propiolactone, formalin, cetyltrimethyl ammonium bromide, triton X-100, and sodium deoxycholate, agents commonly used for virus inactivation and disruption of viral particles during influenza vaccine production. Twenty-two different mycoplasma species (including one to four individual strains of each species) were exposed to these inactivating agents at varying concentrations. The most efficient inactivation of the mycoplasmas evaluated was observed with either 0.5% triton X-100 or 0.5% sodium deoxycholate. Cetyltrimethyl ammonium bromide at concentrations of 0.08% and higher was also able to rapidly inactivate (in less than 30 min) all mycoplasmas tested. In contrast, negligible reduction of mycoplasma titers was observed with 0.0125-0.025% formaldehyde solution. However, increasing the concentration of formaldehyde to 0.1-0.2% improved the mycoplasmacidal effect. Incubation of mycoplasmas with 0.1% beta-propiolactone for 1-24 hours showed a marked mycoplasmacidal effect. A comparison of the mycoplasma inactivation profiles showed that selected species, i.e., M. synoviae, M. gallisepticum, M. orale, M. pneumoniae, and A. laidlawii, represented a set of strains that can be utilized for validating the effectiveness of mycoplasma clearance obtained by the inactivation and viral purification processes used for the manufacture of an inactivated egg-based vaccine.
|Category: Journal Article|
|PubMed ID: #20228111||DOI: 10.1128/AEM.02776-09|
|Includes FDA Authors from Scientific Area(s): Biologics|
|Entry Created: 2011-10-04||Entry Last Modified: 2012-08-29|