20971134
not annotated - annotated - LINNAEUS only
Applied biotechnology for production of immunoglobulin Y specific to hepatitis A virus.
A new protocol for producing polyclonal antibody against hepatitis A virus (HAV) is described. Twenty hens were immunized three times with a commercial HAV vaccine and HAV from a cell culture with three types of adjuvants: CpG oligodeoxynucleotides (CpG-ODN), incomplete Freund's adjuvant and an alum adjuvant. In each of the last two booster inoculations, blood from the birds was collected and tested for HAV antibodies. Egg yolk was separated from egg white and immunoglobulin Y (IgY) antibody was then purified by polyethylene glycol 6000. The mean yield of total protein in yolk was 22.62 mg/mL. Specific activity of the antibody was tested using commercial ELISA, Western blotting, and in vitro neutralization assay demonstrating that anti-HAV IgY bound specifically. After the first immunization, birds immunized with HAV from cell culture plus incomplete Freund's adjuvant with/without CpG-ODN showed highest levels of anti-HAV IgY in serum (p<0.05). Viral combination with CpG-ODN resulted in early response of anti-HAV serum in hens, reflecting the amount of IgY transferred to the egg yolk (p<0.05). The results suggest that egg yolk may be a large scale source of specific antibodies against hepatitis A virus. Further applications of this method have yet to be tested.
Ann file
T1 Species 69 86 hepatitis A virus
N1 Reference T1 Taxonomy:12092
T2 Species 147 164 hepatitis A virus
N2 Reference T2 Taxonomy:12092
T3 Species 166 169 HAV
N3 Reference T3 Taxonomy:12092
T4 Species 192 196 hens
N4 Reference T4 Taxonomy:9031
T5 Species 242 245 HAV
N5 Reference T5 Taxonomy:12092
T6 Species 258 261 HAV
N6 Reference T6 Taxonomy:12092
T7 Species 497 500 HAV
N7 Reference T7 Taxonomy:12092
T8 Out-of-scope 836 839 HAV
N8 Reference T8 Taxonomy:12092
T9 Species 915 918 HAV
N9 Reference T9 Taxonomy:12092
T10 Out-of-scope 1022 1025 HAV
N10 Reference T10 Taxonomy:12092
T11 Out-of-scope 1118 1121 HAV
N11 Reference T11 Taxonomy:12092
T12 Species 1131 1135 hens
N12 Reference T12 Taxonomy:9031
T13 Species 1300 1317 hepatitis A virus
N13 Reference T13 Taxonomy:12092