Production of high-titer bovine rotavirus with trypsin

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Abstract
Titers of bovine rotavirus in excess of 109 immunofluorescent infectious units/ml of culture fluids were produced using trypsin treatment of the virus. Infectivity of preparations of the virus can be increased with as little as 1 mg of trypsin/ml, with maximum increases of 1-2 log10 with 1 .mu.g of trypsin/ml. The virus grows to titers in excess of 108 immunofluorescent units/ml in [Madin-Darby bovine kidney] MDBK, [African green monkey kidney] LLC-MK2, [rhesus monkey kidney] MA-104 and [human cervical carcinoma] HeLa cells. When MDBK cells are infected with a multiplicity of infection of 20, maximum yields of cell-associated, trypsin-enhanceable virus are obtained 4-8 h postinfection. Maximum yields of cell-free, trypsin-enhanceable virus are produced 16-20 h postinfection. Apparently, trypsin can be used to produce high-titer stocks of bovine rotavirus.