Comparative study of magnetic beads and microplates as supports in heterogeneous amplified assay of miRNA-141 by using mismatched catalytic hairpin assembly reaction

Safenkova, I.V.; Burkin, K.M.; Bodulev, O.L.; Razo, S.C.; Ivanov, A.V.; Zherdev, A.V.; Dzantiev, B.B.; Sakharov, I.Y.

Авторы: Safenkova Irina V., Burkin Konstantin M., Bodulev Oleg L., Razo Shyatesa C., Ivanov Aleksandr V., Zherdev Anatoly V., Dzantiev Boris B., Sakharov Ivan Yu
Журнал: Talanta
Том: 247
Год издания: 2022
Издательство: Elsevier BV
Местоположение издательства: Netherlands
Номер статьи: 123535
DOI: 10.1016/j.talanta.2022.123535
Аннотация: Magnetic beads (MBs) are often considered as an effective carrier in heterogeneous assays due to the simplicity ofseparation and washing, and the ability to increase and control the surface area. However, the effect of the MBssurface on the analytical parameters is poorly characterized and is often postulated from intuitive considerations.Herein, experimental evaluation through the comparison of MBs and microwell plate was carried out using themiRNA-141 (biomarker for cancer) as a target, the detection of which was performed by chemiluminescent assaywith a homogeneous mismatched catalytic hairpin assembly (mCHA) reaction. The mCHA reaction produceddouble-stranded (ds) DNA labeled at one end with fluorescein (Flu) for capture with anti-Flu antibodiesimmobilized on a solid carrier, on the other end with biotin for recognition by streptavidin-polyperoxidaseconjugate. The conditions of immobilization of anti-Flu antibody on MBs (a diameter of 440 nm) performedusing a carbodiimide method were optimized by varying the antibody concentration in the reaction solution. Itwas shown that the dependence of chemiluminescent signal as a function of the concentration of anti-FluAb-MBsconjugates had a bell-shaped character. The maximum chemiluminescence was produced at the concentration ofthe conjugates of 2 × 109 particles/mL, with a surface area of 65 mm2. The identical surface area was used uponthe assay performance with polystyrene microplates. Comparison of MBs- and microplate-assays for miRNA-141determination showed that the obtained calibration curves and their detection limit values were the same anddid not depend on the used carrier. The results showed that the choice of a carrier for heterogeneous assaysshould be guided by the convenience of the assay performance, not its surface area.
Добавил в систему: Сахаров Иван Юрьевич

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Comparative study of magnetic beads and microplates as supports in heterogeneous amplified assay of miRNA-141 by using mismatched catalytic hairpin assembly reactionстатья Исследовательская статья

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