Production of Polyclonal Antibodies against the Recombiant Citrus tristeza virus Coat Protein Expressed in Escherichia coli
Production of Polyclonal Antibodies against the Recombiant Citrus tristeza virus Coat Protein Expressed in Escherichia coli
Hala A. Amin1; A. Barakat2; A.A. Abou Zeid1
ABSTRACT
Early diagnosis based on immunoenzymatic techniques requires significant amounts of immunoreagents. Hence, using directional cloning technology, a 38-KDa fusion protein containing a complete coat protein (CP) gene or Citrus tristeza virus (CTV) encoded by the expression vector plasmid (Pinpoint™ Xa3, Promega) with a fragment of biotin binding protein (BBP-rCTV-CP) was highly expressed and purified from E. coli cell culture. Antiserum obtained from rabbits after injection with the BBP-rCTV-CP fusion protein showed comparable reactivity in the serological detection of CTV. This antiserum could be used at dilution or 1: 10.000 at the detection stage in an indirect ELISA (IDAS-ELISA) and was efficient for trapping the virus in standard ELISA. These polyclonal antibodies reacted with a wide range of CTV isolates from different Egyptian geographic sources, and or different biological properties.
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