Электрохимический иммуносенсорный анализ псевдовирусных частиц полиомиелита
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Аннотация
C целью получения псевдовирусных частиц, пригодных для производства вакцин, разработан электрохимический иммуносенсор для оценки сохранности D-антигена штамма Сэбина вируса полиомиелита типа 2 и типа 1 после инактивации. Для иммобилизации моноклональных антител использовали наночастицы золота, получаемые электросинтезом на поверхности рабочего графитового электрода. При увеличении концентрации антигена регистрируется снижение интенсивности сигнала тока пика феррицианида калия, используемого в качестве редокс-зонда. Определены значения пределов обнаружения (LOD) для штаммов Сэбина вируса полиомиелита типов 1 и 2, равные 9.4 DU/мл и 15 DU/мл соответственно (DU/мл - D-антигенные единицы на миллилитр, 1 DU = 15 нг). Рассчитаны константы связывания (Kb) процесса образования комплексов антитело-антиген, 4.68×10⁸ М⁻¹ для Mab2-SII и 1.07×10¹⁰ М⁻¹ для Mab1-SI. Высокоаффинное взаимодействие (Kb ~ 10⁸ М⁻¹ – 10¹⁰ М⁻¹) в системах антитело-антиген свидетельствует о сохранности структуры D-антигена после различных методов инактивации вируса полиомиелита.
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