Abstract
Antigen Used for Immunization
The prokaryotic expression system was used to express the recombinant protein pET-28a/VP7 for immunization.
Method of Immunization
Each mouse was inoculated with 50 μg purified recombinant protein pET-28a (+)/VP7 and equal amount of Complete Freund's adjuvant on day 0. On days 14 and 28, mice were boosted by 50 μg purified recombinant protein pET-28a (+)/VP7 and an equal amount of incomplete Freund's adjuvant.
Parental Cell Line Used for Fusion
SP2/0.
Selection and Cloning Procedure
The positive hybridomas were detected by the recombinant pGEX-6P-1/VP7 using indirect ELISA from well to well. Hybridoma cells from the positive wells were cloned by limiting dilution.
Heavy and Light Chains of Immunoglobulin
IgG.
Specificity
Western blot analysis showed 3H7 reacted with pGEX-6P-1/VP7, pCold-TF/VP7, and pET28a/VP7 against the 25th bluetongue virus and pET28a-VP7 against the 8th bluetongue virus, but not with pET-28a/VP2a, pET-28a/VP2b, pET-28a/VP2c, against the 25th bluetongue virus.
Specific Antigen Identified
The recombinant protein pGEX-6P-1-BTV25-VP7, pCold-TF-BTV25-VP7, pET28a-BTV25-VP7, pET-28a-BTV25-VP2a, pET-28a-BTV25-/VP2b, pET-28a-BTV-VP2c, and pET28a- BTV8-VP7.