For the first immunization, the immunogen
(0.1 mg) was suspended in 2 ml of 0.5× phosphate buffer saline
(PBS), emulsified with 2 ml Freund’s complete adjuvant and
injected subcutaneously in two rabbits for each immunogen.
Four booster dose immunizations were given at monthly
intervals after the first immunization using 0.1 mg TSWV virions
in 2 ml of 0.5× PBS 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 ProteinA affinity chromatography
columns and dialyzed against 10 mM sodium phosphate
buffer (pH 7.2), according to manufacturer’s instructions (PALL,
USA).