Preparation and properties evaluation of the recombinant analogue of birch pollen allergen Bet v 1



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Abstract

Background. For wide introduction of the component allergy diagnostics in laboratory practice it is necessary to create methods of production, purification and verification of synthetic allergens. Major birch pollen protein Bet v 1, a representative of the superfamily PR-10, is interesting due to the high incidence of sensitization and the manifestation of cross-reactive properties. Methods. To receive a synthetic protein DNA cloning techniques were used, followed by expression in E. coli. Purification was made by metal-affinity chromatography. Evaluation of immunochemical properties was held in several tests including ELISA analysis and line-blot. Results. We have obtained high-yield bacterial strain E. coli, containing expression vector for protein Bet v 1 production. Physico-chemical and immunological properties were estimated in various test systems with the reference panel of serum. Conclusion. It was shown that the resulting synthetic analogue of protein Bet v 1 can be used to measure the presence of specific immunoglobulin E in serum samples. This allergenic component may be used for estimation of each component of the sensitization profile.

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About the authors

A E Pavlov

Vega Ltd. group of Companies «Alkor Bio»; Saint-Petersburg State University

Email: apavlov@alkorbio.ru
Saint-Petersburg, Russia

N A Seylieva

Vega Ltd. group of Companies «Alkor Bio»

Saint-Petersburg, Russia

O YU Mukhortykh

Vega Ltd. group of Companies «Alkor Bio»

Saint-Petersburg, Russia

V E Stefanov

Saint-Petersburg State University

Saint-Petersburg, Russia

References

  1. Madsen C. Prevalence of food allergy: an overview. Proc. Nutr. Soc. 2005, v. 64, p. 413-417.
  2. Ebo D.G., Hagendorens M.M., Bridts C.H., Stevens W.J. In vitro diagnosis of IgE-mediated allergy: breakthroughs in the last decade. Expert. Rev. Clin. Immunol. 2012, v. 8, p. 9-11.
  3. Eriksson N.E., Ahlstedt S. Diagnosis of reaginic allergy with house dust, animal dander and pollen allergens in adult patients. V. A comparison between the enzyme-linked immunosorbent assay (ELISA), provocation tests, skin tests and RAST Int. Arch. Allergy Appl. Immunol. 1977, v 54, p. 88-95.
  4. Dolen W.K. Allergen extract standartization: reality, myth, or dream? Ann. Allergy Asthma Immunol. 1995, v. 75, p. 81-82.
  5. Ipsen H., Løwenstein H. Isolation and immunochemical characterization of the major allergen of birch pollen (Betula verrucosa). J. Allergy Clin. Immunol. 1983, v. 72, p. 150-159.
  6. Павлов А.Е., Сейлиева Н.А., Стефанов В.Е. Перспективы развития молекулярной диагностики аллергии в формате микрочипа. Клин. лаб. диагн. 2011, № 12, с. 3-7.
  7. Asturias J.A., Ibarrola I., Amat P. et al. Purified allergens vs. complete extract in the diagnosis of plane tree pollen allergy. Clin. Exp. Allergy. 2006, v. 36, p. 1505-1512.
  8. Midoro-Horiuti T., Brooks E.G., Goldblum R.M. Pathogenesis-related proteins of plants as allergens. Ann. Allergy Asthma Immunol. 2001, v. 87, p. 261-271.
  9. WHO/IUIS Allergen Nomenclature Sub-committee: [www. allergen.org]. URL: http://www.allergen.org/.
  10. Swoboda I., Jilek A., Ferreira F. et al. Isoforms of Bet v 1, the major birch pollen allergen, analyzed by liquid chromatography, mass spectrometry and cDNA cloning. J. Biol. Chem. 1995, v. 270, p. 2607-2613.
  11. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970, Aug 15, v. 227 (5259), р. 680-685.

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