Ukr.Biochem.J. 2025; Volume 97, Issue 4, Jul-Aug, pp. 102-109

doi: https://doi.org/10.15407/ubj97.04.102

Human ADA2-specific IgY antibodies from hen eggs with tolerance to human IgG

10.09.2025   Uncategorized   No comments

E. Yu. Sarkisyan1, H. A. Haroutyunyan2, Ye. G. Sargisova1*

1H. Buniatian Institute of Biochemistry of National Academy of Sciences of Armenia, Yerevan;
2Laboratory of Biochemical and Biophysical Investigations,
Scientific-Research Centre, Yerevan State Medical University after Mkhitar Heratsi, Armenia;
*e-mail: lizasar2001@yahoo.com

Received: 22 April 2025; Revised: 09 July 2025;
Accepted: September 2025; Available on-line: 2025

Purification of human plasma adenosine deaminase 2 (ADA2) is challenging due to its low physiological concentration and co-purification of human immunoglobulin G (IgG), which has similar physicochemical properties. This study describes a novel approach exploiting avian immune tolerance to human IgG to selectively produce antibodies against native and active human ADA2. Chickens were rendered tolerant to human IgG, enabling subsequent immunization with a preparation containing both ADA2 and human IgG. This strategy successfully elicited a specific immune response, yielding antibodies exclusively to ADA2. These anti-ADA2 antibodies (IgY), purified from egg yolks, were subsequently employed in an affinity column. This immunopurification method allowed the isolation of an electrophoretically homogeneous preparation of human ADA2. This study demonstrates the utility of inducing immune tolerance to human IgG in chickens.

Keywords: , , , , ,

References:

  1. Brent L. Transplantation tolerance – a historical introduction. Immunology. 2016;147(3):267-268. PubMed, PubMed, CrossRef
  2. Ribatti D. Peter Brian Medawar and the discovery of acquired immunological tolerance. Immunol Lett. 2015;167(2):63-66. PubMed, PubMed, CrossRef
  3. Luo X, Miller SD, Shea LD. Immune Tolerance for Autoimmune Disease and Cell Transplantation. Annu Rev Biomed Eng. 2016;18:181-205. PubMed, PubMed, CrossRef
  4. Akdis CA. Therapies for allergic inflammation: refining strategies to induce tolerance. Nat Med. 2012;18(5):736-749. PubMed, CrossRef
  5. Wolfe HR, Tempelis C, Mueller A, Reibel S. Precipitin production in chickens. XVIL The effect of massive injections of bovine serum albumin at hatching on subsequent antibody production. J Immunol. 1957;79(2):147-153. PubMed
  6. Stevens KM, Pietryk HC, Ciminera JK. Acquired immunological tolerance to a protein antigen in chickens. Br J Exp Pathol. 1958;39(1):1-7. PubMed, PubMed
  7. Ivanyi J, Hraba T. A contribution to the study of immunological tolerance to protein antigen in the chicken. Folia Biol (Praha). 1963;9:354-363. PubMed
  8. Zhao C, Song C, Wang X, Li Z, Sha J, Han H, Zhang Y, Li Z. Induction of immunological tolerance in chickens inoculated with xenogeneic antigens at an early stage of embryonic development. Dev Comp Immunol. 2006;30(4):431-440. PubMed, CrossRef
  9. Antonioli L, Colucci R, La Motta C, Tuccori M, Awwad O, Da Settimo F, Blandizzi C, Fornai M. Adenosine deaminase in the modulation of immune system and its potential as a novel target for treatment of inflammatory disorders. Curr Drug Targets. 2012;13(6):842-862. PubMed, CrossRef
  10. Palomino JC. Nonconventional and new methods in the diagnosis of tuberculosis: feasibility and applicability in the field. Eur Respir J. 2005;26(2):339-350. PubMed, CrossRef
  11. Andreasyan NA, Hairapetian HL, Sargisova YG, Mardanyan SS, Badalyan LT, Khanoyan AS. Activity of adenosine deaminase and its isoforms in pleural fluid in tuberculous pleuritis. Med Sci Monit. 2002;8(10):CR708-CR712. PubMed
  12. Dos Santos Jaques JA, Becker LV, Souza Vdo C, Leal CA, Bertoldo TM, de Vargas Pinheiro K, Morsch VM, Schetinger MR, Leal DB. Activities of enzymes that hydrolyze adenine nucleotides in lymphocytes from patients with rheumatoid arthritis. Cell Biochem Funct. 2013;31(5):395-399. PubMed, CrossRef
  13. Lee PY. Vasculopathy, Immunodeficiency, and Bone Marrow Failure: The Intriguing Syndrome Caused by Deficiency of Adenosine Deaminase 2. Front Pediatr. 2018;6:282. PubMed, PubMed, CrossRef
  14. Caorsi R, Penco F, Schena F, Gattorno M. Monogenic polyarteritis: the lesson of ADA2 deficiency. Pediatr Rheumatol Online J. 2016;14(1):51. PubMed, PubMed, CrossRef
  15.  Zavialov AV, Gracia E, Glaichenhaus N, Franco R, Zavialov AV, Lauvau G. Human adenosine deaminase 2 induces differentiation of monocytes into macrophages and stimulates proliferation of T helper cells and macrophages. J Leukoc Biol. 2010;88(2):279-290. PubMed, PubMed, CrossRef
  16. Greiner-Tollersrud OK, Boehler V, Bartok E, Krausz M, Polyzou A, Schepp J, Seidl M, Olsen JO, Smulski CR, Raieli S, Hübscher K, Trompouki E, Link R, Ebersbach H, Srinivas H, Marchant M, Staab D, Guerini D, Baasch S, Henneke P, Kochs G, Hartmann G, Geiger R, Grimbacher B, Warncke M, Proietti M. ADA2 is a lysosomal DNAse regulating the type-I interferon response. bioRxiv. 2020;2020-06. CrossRef
  17. Greiner-Tollersrud OK, Krausz M, Boehler V, Polyzou A, Seidl M, Spahiu A, Abdullah Z, Andryka-Cegielski K, Dominick FI, Huebscher K, Goschin A, Smulski CR, Trompouki E, Link R, Ebersbach H, Srinivas H, Marchant M, Sogkas G, Staab D, Vågbø C, Guerini D, Baasch S, Latz E, Hartmann G, Henneke P, Geiger R, Peng XP, Grimbacher B, Bartok E, Alseth I, Warncke M, Proietti M. ADA2 is a lysosomal deoxyadenosine deaminase acting on DNA involved in regulating TLR9-mediated immune sensing of DNA. Cell Rep. 2024;43(11):114899. PubMed, CrossRef
  18. Kim SH, Park MK, Kim JY, Chuong PD, Lee YS, Yoon BS, Hwang KK, Lim YK. Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies. J Vet Sci. 2005;6(1):41-46. PubMed, CrossRef
  19. Brujeni GN, Gharibi D. Development of DNA-designed avian IgY antibodies for detection of Mycobacterium avium subsp. paratuberculosis heat shock protein 70 (Hsp70) and anti-Hsp70 antibodies in the serum of normal cattle. Appl Biochem Biotechnol. 2012;167(1):14-23. PubMed, CrossRef
  20. Łupicka-Słowik A, Psurski M, Grzywa R, Bobrek K, Smok P, Walczak M, Gaweł A, Stefaniak T, Oleksyszyn J, Sieńczyk M. Development of Adenosine Deaminase-Specific IgY Antibodies: Diagnostic and Inhibitory Application. Appl Biochem Biotechnol. 2018;184(4):1358-1374. PubMed, PubMed, CrossRef
  21. Andreasyan NA, Hairapetyan HL, Sargisova YG, Mardanyan SS. ADA2 isoform of adenosine deaminase from pleural fluid. FEBS Lett. 2005;579(3):643-647. PubMed, CrossRef
  22. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-685. PubMed, CrossRef
  23. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72(1-2):248-254. PubMed, CrossRef
  24. Ouchterlony O. Diffusion-in-gel methods for immunological analysis. Prog Allergy. 1958;5:1-78. PubMed, CrossRef
  25. Polson A, Coetzer T, Kruger J, von Maltzahn E, van der Merwe KJ. Improvements in the isolation of IgY from the yolks of eggs laid by immunized hens. Immunol Invest. 1985;14(4):323-327. PubMed, CrossRef
  26. Yeritsyan HE, Gasparyan VK. Homogeneous immunoassay for human IgG using oriented hen egg IgY immobilized on gold sol nanoparticles. Microchim Acta. 2012;176:117-122. CrossRef
  27. Pham NB, Meng WS. Protein aggregation and immunogenicity of biotherapeutics. Int J Pharm. 2020;585:119523. PubMed, PubMed, CrossRef
  28. Lundahl MLE, Fogli S, Colavita PE, Scanlan EM. Aggregation of protein therapeutics enhances their immunogenicity: causes and mitigation strategies. RSC Chem Biol. 2021;2(4):1004-1020. PubMed, PubMed, CrossRef
  29. Lee L, Samardzic K, Wallach M, Frumkin LR, Mochly-Rosen D. Immunoglobulin Y for Potential Diagnostic and Therapeutic Applications in Infectious Diseases. Front Immunol. 2021;12:696003. PubMed, PubMed, CrossRef
  30. Vieira-Pires RS, Morgan PM, Choraria A, Gandhimathi C, Michael A, Ôchoa-Pires T, Salimian J, Rajan S, Subramani M. Applications of IgY in Human Medicine. In: Zhang XY, Vieira-Pires RS, Morgan PM, Schade R. (eds) IgY-Technology: Production and Application of Egg Yolk Antibodies. Springer, Cham., 2021:237-278. CrossRef
Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License.