Assay for porcine circovirus production

Chemistry: molecular biology and microbiology – Virus or bacteriophage – except for viral vector or...

Reexamination Certificate

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C435S325000

Reexamination Certificate

active

07358075

ABSTRACT:
The present invention provides methods for the determination of the viral titer of a culture of host animal host cells infected with a circovirus. The FACS-based methods of the invention may include determining the viability of the host cells in a cell culture medium supernatant and of those cells that remain adherent to a solid support. Detecting and measuring the percentage of cells that expressed the viral antigens ORF1 and ORF2 may determine the viral load of the cultured host cells. The yield of the virus may be established by the detection and measurement of both antigens in supernatant cells, for example 5 to 7 days from when the host cells are transferred to a serum free medium. The methods of the invention may yield rapid quantitative data. This allows the repeated in-process monitoring of the viral production throughout the incubation period, and ready selection of the most appropriate harvesting point.

REFERENCES:
Liu et al. Virology, 2001, vol. 285, pp. 91-99.
Gilpin et al. Veterinary Immunology and Immunopathology, 2003, vol. 94, pp. 149-161.
Meerts et al. Archives of Virology, Mar. 2005, vol. 150, pp. 427-441.
McNeilly, F., et al.: Production, Characterisation and Applications of Monoclonal Antibodies to Procine Circovirus 2, Archives of Virology, New York, NY, vol. 146, No. 5, 2001, p. 909-922.

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