Purification and characterization protein of anti-dengue specific immunoglobulin Y for diagnostic kit of dengue

This study aimed to produce immunoglobulin Y (IgY) specific to dengue virus which could be used for diagnostic kit of dengue. Lohman laying hens were immunized intramuscularly with antigenic of dengue. Egg yolk was separated from egg white and IgY antibody was then purified by multiple polyethylene glycol (PEG) 6000 extraction and ammonium sulfate purification steps. The IgY concentration in egg yolk increased during the immunization period until week 6 where it began to increase dramatically at 2 weeks and it reached a plateau at 4 weeks after immunization. After week 6 the levels decreased gradually. Antibody of dengue was detected and produce a specific line of precipitation in agar gel precipitation test (AGPT) beginning the second week after the first immunization. Analysis of results obtained with ELISA showed significant increase in the dengue-specific antibodies after two weeks from the primary immunization. Through the effect of boostering; the anti-dengue antibody levels reached a plateau at four weeks from the primary immunization and remained significantly higher till the end of observation period. SDS-PAGE revealed the IgY preparation to be pure and dissociated into protein bands with molecular weights of 145; 66; 45, 33 and 26 kDa and western blot analysis revealed the presence of anti-dengue IgY in egg yolks protein, with a molecular weights of approximately 66 kDa. These results suggested that chicken IgY could be a practical strategy in large-scale production of specific anti-dengue Ig Y for diagnostic KIT of dengue.