Production of anti-idiotype immunoglobulin Y against African swine fever virus using purified porcine immunoglobulin G

Auliya Rahma Riana; I Kadek Mahendra Putra; I Wayan Teguh Wibawan; Okti Nadia Poetri

doi:10.29244/currbiomed.4.2.72

Authors

Auliya Rahma Riana Study Program of Veterinary Medicine, School of Veterinary Medicine and Biomedical Science, IPB University, Bogor, Indonesia https://orcid.org/0009-0001-8283-4732
I Kadek Mahendra Putra Study Program of Veterinary Medicine, School of Veterinary Medicine and Biomedical Science, IPB University, Bogor, Indonesia https://orcid.org/0009-0003-3664-0608
I Wayan Teguh Wibawan Division of Medical Microbiology, School of Veterinary Medicine and Biomedical Science, IPB University, Bogor, Indonesia https://orcid.org/0000-0003-3165-080X
Okti Nadia Poetri Division of Medical Microbiology, School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia https://orcid.org/0000-0002-8028-6023

DOI:

https://doi.org/10.29244/currbiomed.4.2.72

Keywords:

African swine fever virus, anti-idiotype antibody, immunoglobulin G, immunoglobulin Y, surrogate antigen

Abstract

Background African swine fever (ASF) is a lethal viral disease threatening the global swine industry. Anti-idiotype antibodies (Ab2) mimicking viral epitopes serve as promising surrogate antigens for vaccine and diagnostic development.

Objective This study aimed to produce anti-idiotype immunoglobulin Y (IgY) against African swine fever virus (ASFV) by immunizing laying hens with purified ASFV-specific porcine immunoglobulin G (IgG).

Methods ASFV-specific IgG (Ab1) was purified from ASF-vaccinated pigs serum and convalescent plasma of naturally infected pigs by stepwise ammonium sulfate precipitation (25% and 50% saturation) and dialysis. Purified IgG was characterized using Bradford assay, indirect ELISA, and SDS-PAGE. Two Hy-Line Brown laying hens were immunized with serum-derived (A1) or plasma-derived (A2) IgG, using Freund's complete adjuvant for primary immunization, followed by three booster injections with Freund's incomplete adjuvant at two-week intervals. Serum and egg yolk samples collected over 13 weeks were analyzed using agar gel precipitation test (AGPT).

Results Purification increased total protein while retaining ASFV-specific antibody reactivity. SDS-PAGE confirmed porcine IgG heavy (~53–55 kDa) and light (~24–25 kDa) chains, alongside additional bands indicating partial purification. Serum antibodies reactive to porcine IgG were first detected at weeks 2 (A2) and 4 (A1) post-primary immunization, with egg yolk antibodies appearing one week later. Both serum and egg yolk achieved a maximum AGPT titer of 3 log₂ (1:8 dilution).

Conclusion Immunization with ASFV-specific porcine IgG successfully elicited reactive IgY in laying hens. Further validation of anti-idiotype specificity and antigen-mimicking properties is required before considering these antibodies as surrogate antigens for ASF vaccine and diagnostic development.