Abstract
A method was developed for the production of Eastern equine encephalomyelitis vaccine from virus grown in rolling-bottle cultures (840 cm2 growth area) of chick embryo cells. The PE-6 strain of virus was propagated in chick embryo cell roller cultures maintained on serum-free medium 199 containing 0.25% human serum albumin and antibiotics (MM). A multiplicity of inoculum of 0.005 yielded acceptable titers of virus at a convenient harvest time of 18 to 24 hr and reduced the carry-over of extraneous material from the virus seed. Growth studies in which 100, 200, or 300 ml of MM was used showed that use of 300 ml of MM offered two advantages: (i) cytopathic effects were less at the 18- to 24-hr harvest time, thereby decreasing cellular material in the final product, and (ii) total virus yield was not substantially reduced, thus permitting large-scale production of virus for further processing. Studies on formalin inactivation at 37 C indicated that the virus was inactivated by 0.05% formalin within 12 to 16 hr and with 0.1% formalin within 6 to 8 hr. Antigen extinction tests in hamsters revealed excellent potency (e.g., median-effective-dose values of 0.069 to 0.012 ml) for both fluid and freeze-dried products. The advantages of the roller-bottle technique in vaccine production are discussed.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bartelloni P. J., McKinney R. W., Duffy T. P., Cole F. E., Jr An inactivated eastern equine encephalomyelitis vaccine propagated in chick-embryo cell culture. II. Clinical and serologic responses in man. Am J Trop Med Hyg. 1970 Jan;19(1):123–126. doi: 10.4269/ajtmh.1970.19.123. [DOI] [PubMed] [Google Scholar]
- Cole F. E., Jr, McKinney R. W. Use of hamsters of potency assay of Eastern and Western equine encephalitis vaccines. Appl Microbiol. 1969 Jun;17(6):927–928. doi: 10.1128/am.17.6.927-928.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maire L. F., 3rd, McKinney R. W., Cole F. E., Jr An inactivated eastern equine encephalomyelitis vaccine propagated in chick-embryo cell culture. I. Production and testing. Am J Trop Med Hyg. 1970 Jan;19(1):119–122. doi: 10.4269/ajtmh.1970.19.119. [DOI] [PubMed] [Google Scholar]
- Robinson D. M., Berman S., Lowenthal J. P., Hetrick F. M. Western equine encephalomyelitis vaccine produced in chick embryo cell cultures. Appl Microbiol. 1966 Nov;14(6):1011–1014. doi: 10.1128/am.14.6.1011-1014.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]