Purification of hop mosaic virus and preparation of antiserum suitable for ELISA
1993
Polak, J. | Chod, J. | Pivalova, J. | Jokes, M. (Vyzkumny Ustav Rostlinne Vyroby, Prague-Ruzyne (Czech Republic))
Specific antibodies against hop mosaic virus (HMV) were prepared. HMV was multiplied in plants of tobacco Nicotiana clevelandii x Nicotiana glutinosa. Homogenization of infected plants and purification of HMV was done in 0.05M borate buffer pH 8.0. The virus was precipitated by 5 % polyethyleneglycol 6000 from sap of infected plants and purified by differential centrifugation and centrifugation on sucrose gradient (10-40 %). Virus fraction was obtained on ISCO fractionator and the rest of normal proteins was removed by centrifugation on 20 % sucrose cushion. HMV suspension contained filamentous particles 650 nm long. HMV suspension was used for immunization of rabbits.Titre of antisera obtained was 1:512. Immunoglobulins of HMV were isolated and conjugated with alkaline phosphatase, HMV was detected with these antibodies in infected plants of tobacco and hop.
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