For the first immunization,the immunogen (0.1 mg) was suspended in 2 ml of 0.5X
phosphate buffer saline (PBS), emulsified with 2 ml Fre-und’s complete adjuvant and injected subcutaneously in
two rabbits for each immunogen. Four booster dose immu-nizations were given at monthly intervals after the first
immunization using 0.1mg TBSV virions in 2 ml of 0.5XPBS buffer emulsified with Freund’s incomplete adjuvant.
About 10 - 12 ml serum was collected from the marginal ear vein, 14 days after each booster. Sera that exhibited a
strong immunogenic response were further purified by affinity chromatography using Protein A affinity chroma-
tography columns and dialyzed against 0.01M, sodium phosphate buffer (pH7.2), according to manufacturer’s
instructions (PALL, USA).