Preparation High Titer Anti-serum of Porcine Circovirus Type Ⅱ Capsid Protein by Hydrodynamics Gentic Immunization
1, 2, 3, 4
1. Hebei Agricultural University; Hebei University of engineering 2. Hebei Normal University of Science and Technology, 3. Hebei Agricultural University 4. Hebei University of engineering
Abstract In order to establish a simple and efficient anti-serum preparation method for molecular biology research, in this research, porcine circovirus type Ⅱ capsid protein gene was selected to research on the possiblity of preparing high titer anti-serum by hydrodynamics gentic immunization. Endotoxin free plasmid of pcDNA-Cap, with would express porcine circovirus type Ⅱ capsid protein in the exkaryotic cell, was prepared using endotoxin free plasmid preparing kit and was delivered by hydrodynamics tail vein injection method for genetic immunization of mice(Mus musculus). After 5 times continuous immunization, the blood serum was collected. Using porcine circovirus type Ⅱ capsid protein which has been deleted its nuclear laction signal sequence at its N-terminal and expressed in Escherichia coli as antigen, the prepared anti-serum was tested by enzymeliked immuno sorbent assay(ELISA) and Western blot. The results showed that the anti-serum prepared by hydrodynamics tail vein gene delivery method had very high titer(1∶1 000 000) by ELISA and could detect at least 10 ng antigen after 5 000-fold dilution in Western blotting analysis. These results showed that hydrodynamics gene delivery method may become an efficient method for preparing antibody which will be used in porcine circovirus type Ⅱ related research. This study provides an alternative method for prevention the disease caused by porcine circovirus type Ⅱ.
