Abstract
The use of egg yolk antibodies—IgY technology—represents an alternative to the production of mammalian immunoglobulins and has several advantages regarding animal welfare and lower costs of production. The use of adjuvants to achieve the hyperimmunization of laying hens plays a key role in the success of the production of high levels of the antibodies. In the present work, two different adjuvant systems (Freund’s adjuvants and MontanideTM ISA 71 VG) were compared to produce IgY anti-Bothrops alternatus. For the first immunization, formalin-inactivated Salmonella was added to MontanideTM ISA 71 VG to emulate Freund’s complete adjuvant which includes a mycobacteria antigen. After eight immunizations, IgY produced by using either adjuvant was able to neutralize the lethal activity of the venom in a mouse model, but differences were found regarding the recognition of components of the venom between the two adjuvants tested. Overall, MontanideTM adjuvant used in this work could be a good alternative choice to produce antibodies capable of neutralizing the lethality of complex antigens. This adjuvant is commercially available and used in the formulation of several poultry vaccines and could be used for the IgY technology instead of traditional immunomodulators such as Freund’s adjuvants.
Key points
• IgY extracts recognized major components of the venom.
• Avidity indexes of the IgY extracts increased after the successive immunizations.
• IgY obtained by two adjuvant systems neutralized the lethal activity of the venom.
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Data availability
The data that support the findings of this study are available from the corresponding author, CLL, upon reasonable request.
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Acknowledgements
We would like to thank Dr. Christian Dokmetjian and Dr. Matias Fingermann for providing the venoms, and Mr. Alejandro Vivas and Esteban Medina for their help in the maintenance of the animals.
Funding
This work was supported by the Project FITS Salud PPM03 from the Fondo Argentino Sectorial (FONARSEC) of the National Agency for Science and Technology Promotion-MinCyT of Argentina.
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CLL and PC conceived and designed the research. CLL, AC, VM, and AC conducted the experiments. PJ and PG contributed with new reagents. CLL analyzed the data. CLL and PC wrote the manuscript. PC, MFM, and PG obtained funding for the project. All of the authors read and approved the manuscript.
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This study meets the ARRIVE guidelines (du Sert et al. 2020), and the experiments were approved by the Institutional Animal Care and Use Committee from the Centro de Investigación en Ciencias Veterinarias y Agronómicas of the Instituto Nacional de Tecnología Agropecuaria (CICVyA-INTA), Argentina (Procedure #43/2018).
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Leiva, C.L., Cangelosi, A., Mariconda, V. et al. Use of adjuvant ISA VG 71 to produce neutralizing egg yolk antibodies against bothropic venom. Appl Microbiol Biotechnol 107, 1947–1957 (2023). https://doi.org/10.1007/s00253-023-12409-3
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DOI: https://doi.org/10.1007/s00253-023-12409-3