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14 August 2012

Preparation and properties evaluation of the recombinant analogue of birch pollen allergen Betv1

Pavlov A.E. 1,2, Seylieva N.A. 1, Mukhortykh O.Yu. 1, Stefanov V.E. 2

1. Vega Ltd. Group of Companies «Alkor Bio», Saint-Petersburg;

2. Saint-Petersburg State University, Saint-Petersburg. 

Keywords: allergy diagnostics, birch pollen, recombinant allergen, Betv1.

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 Betv1, 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 were held in several test including ELISA analysis and line-blot.

Results. We have obtained high-yield bacterial strain E.coli, containing expression vector for protein Betv1 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 Betv1 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.