Diagnostics of Allergies: Realities and Prospects. Part 2

Khokha, R. N.

doi:10.25298/2221-8785-2020-18-4-481-486

Cited by 2 publications

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“…Поэтому диагностика с применением экстрактов позволяет определить только источники аллергенов, но не отдельные молекулы. В аллерген-специфической иммунотерапии (АСИТ) следствием невозможности прогнозирования способности таких аллергенов индуцировать иммунологическую толерантность становится большая вариативность результатов терапии и недостаточная приверженность лечению [4][5][6].…”

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Obtaining Recombinant Polypeptide Bet v 1, the Main Allergen of Birch Pollen, and Creation of an Experimental Test System for Specific IgE Antibodies Identifying on Its Basis

Пархомчук,

Фомина,

Григорьева

et al. 2024

Лабораторная Диагностика. Восточная Европа

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Введение. Во всем мире наблюдается рост числа аллергических заболеваний. Внедрение рекомбинантных технологий в аллергологию позволило определить белковые компоненты, отвечающие за формирование аллергической реакции. Синтез рекомбинантных аллергенов дает возможность целенаправленно получать пептиды с заданными свойствами. Цель. Получить рекомбинантный полипептид Bet v 1.0101 в прокариотических клетках Escherichia coli, штамм BL21 (DE3). Материалы и методы. В качестве матрицы использована тотальная РНК пыльцы березы повислой, собранная на территории Республики Беларусь. Для получения целевого полипептида использована прокариотическая система Escherichia coli, штамм BL21(DE3), и экспрессирующий вектор pJC40. Очистка проведена методом металл-хелатной хроматографии в денатурирующих условиях. Для подтверждения специфичности белка применен разработанный иммуноферментный лиа-тест, который основан на реакции агглютинации («антиген – антитело»), где в качестве антигена выступает сорбированный на нитроцеллюлозной мембране полипептид Bet v 1.0101. Результаты. Получен высокоочищенный рекомбинантный полипептид Веt v 1 (изоформа Bet v 1.0101), не содержащий примесей бактериальных белков, обладающий антигенными свойствами. Показана возможность использования белка для выявления специфических антител в иммуноферментном лиа-тесте с референс-сыворотками. Заключение. Открывается возможность использования полученного рекомбинантного полипептида для диагностики аллергических заболеваний и в аллергенспецифической иммунотерапии. Introduction. Allergic diseases are on the rise worldwide. The introduction of recombinant technologies in allergology has allowed identifying protein components responsible for allergic reaction formation. Recombinant allergens synthesis makes it possible to purposefully obtain peptides with targeted properties. Purpose. To obtain recombinant polypeptide Bet v 1.0101 in prokaryotic cells of Escherichia coli, strain BL21 (DE3). Materials and methods. The total RNA of birch pollen collected on the territory of the Republic of Belarus was used as a matrix. To obtain the target polypeptide, the prokaryotic Escherichia coli system, strain BL21(DE3), and the expression vector pJC40 were used. The purification was carried out by metal-chelate chromatography under denaturing conditions. To confirm the protein specificity, an elaborated enzyme immunoassay was used based on agglutination reaction ("antigen – antibody"), where the polypeptide Bet v 1.0101 sorbed on a nitrocellulose membrane acted as antigen. Results. A highly purified recombinant polypeptide Bet v 1 (isoform Bet v 1.0101), not containing bacterial protein impurities and possessing antigenic properties, was obtained. The feasibility of using the protein for specific antibodies detecting by enzyme immunoassay with reference sera has been shown. Conclusion. The opportunity of using the obtained recombinant polypeptide in allergic conditions diagnostics and in allergen-specific immunotherapy has been discovered.

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Obtaining Recombinant Polypeptide Bet v 1, the Main Allergen of Birch Pollen, and Creation of an Experimental Test System for Specific IgE Antibodies Identifying on Its Basis

Пархомчук,

Фомина,

Григорьева

et al. 2024

Лабораторная Диагностика. Восточная Европа

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The spectrum of sensitization in children with respiratory allergic diseases living in the Rostov region, according to molecular allergodiagnostics

Semernik,

Lebedenko,

Sarychev

et al. 2024

Russian Journal of Allergy

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BACKGROUND: Due to the widespread occurrence of respiratory allergic diseases, it is of particular interest to study the regional spectrum of sensitization of the population using a modern method-molecular allergodiagnostics. OBJECTIVE: to determine the spectrum of sensitization in children with allergies living in the Rostov region by the method of molecular allergodiagnostics. METHODS: An open multicenter prospective controlled non-randomized study included 67 children with respiratory allergic diseases living in the Rostov region. The patients were examined using the Allergy Explorer 2 (ALEX2) allergy chip. Statistical processing of the results was carried out using the Microsoft Office Excel 2003 and Statistica 12.0 for Windows software package. RESULTS: According to the conducted studies, it was found that the most common sensitization in children of the Rostov region is caused by weed pollen, pet allergens, pollen of trees, cereals, fruits, mold and yeast fungi, class 4 sensitization was also noted to uteroglobin and lipocalin. Molecular analysis showed that an allergic reaction to ragweed pollen prevailed among pollen allergens - the allergocomponent Amb a1 caused sensitization in 27 (40.3%) children, and nettle (Urt d) - 23 (34.33%) of the examined. The analysis of the allergen components of tree pollen showed that the most common positive reaction was to date palm (22.39%), cryptomeria japonica (19.4%), while only 6 (8.96%) patients had a "true" allergy to birch pollen (positive component Bet v 1). If we talk about sensitization to house dust mites and mold allergens, it should be noted that the molecules Alternaria alternata (20.89%), Blomia tropicalis (23.88%) and Glycyphagus domesticus (23.88%) prevailed in the spectrum of this group of allergens. CONCLUSION. In the course of the study, it was found that most often in children of the Rostov region with respiratory allergic diseases, sensitization to weed pollen was determined. This study allowed not only to determine the class of sensitization, but also to identify the allergen components that play a leading role in the development of pathology.

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Diagnostics of Allergies: Realities and Prospects. Part 2

Cited by 2 publications

References 0 publications

Obtaining Recombinant Polypeptide Bet v 1, the Main Allergen of Birch Pollen, and Creation of an Experimental Test System for Specific IgE Antibodies Identifying on Its Basis

Obtaining Recombinant Polypeptide Bet v 1, the Main Allergen of Birch Pollen, and Creation of an Experimental Test System for Specific IgE Antibodies Identifying on Its Basis

The spectrum of sensitization in children with respiratory allergic diseases living in the Rostov region, according to molecular allergodiagnostics

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