ANTI-miRNA IMMOBILIZATION OPTIMIZATION ON THE SCREEN PRINTED ELECTRODES FOR ELECTROCHEMICAL miRNA BIOSENSORS

Yıl 2022, , 1 - 10, 19.01.2022

Karima Sahtani Yakup Aykut , Nilay Aladağ Tanik

https://doi.org/10.18036/estubtdc.866279

Öz

Synthetically produced miRNA molecules plays an important role as biomarker to examine and investigate the diagnosis of some diseases including cancer. In order to develop a sensitive electrochemical biosensor system for the detection of miRNA molecules, the anti-miRNA molecules are synthesized and immobilized on the biosensor surfaces and observe the signal changes via a proper measurement. Immobilization time and temperature along with the anti-miRNA concentration are critically important for an appropriate observation of the miRNA detection sensitivity of the prepared biosensor system. In this regard, synthetically produced anti-miRNA (anti-miR451(G)) was purchased and diluted into different concentration by using phosphate buffer solution. Then, the solutions were immobilized on the screen printed electrodes (SPEs) and the guanine oxidation signal of the anti-miRNA molecules were observed via differential pulse voltammetry method (DPV). An appropriate concentration of the solution was selected and dropped on the SPEs and held on at different temperatures (-18, +5 and +25 oC) for 1, 3, 14 and 21 days and DPV measurements were conducted to investigate the optimum immobilization time and temperature. The result shown that guanine oxidation signal was increased by increasing the concentration of the genetic molecules in the immobilization solution and increased less after that point when the concentration increased more because the surface reached to a certain saturation value . The guanine oxidation signal revealed that the best suitable storing temperature after the immobilization was +5 oC determined.

Anahtar Kelimeler

miRNA detection, Screen printed electrode, DPV measurement, Guanine oxidation

Destekleyen Kurum

The Scientific and Technological Research Council of Turkey (TUBITAK)

Proje Numarası

118M231

Teşekkür

The present study was part of a research project funded by The Scientific & Technological Research Council of Turkey (TUBITAK, Project no. 118M231).

ELEKTROKİMYASAL miRNA BİYOSENSÖRLER İÇİN ANTİ-miRNA MOLEKÜLÜNÜN YÜZEY-BASKI ELEKTROTLAR ÜZERİNE İMMOBİLİZASYONUNUN OPTİMİZASYONU

Öz

miR-1841 geni insan genomunda bulunan ve bazı kanser türlerinin erken tespiti için biyobelirteç olduğu literatürde belirtilmektedir. Genetik moleküllerin rollerinin kavranmasında sentetik olarak üretilebilen miRNA molekülleri ile yapılan çalışmalar önemli yer tutmaktadır. miRNA moleküllerinin elektrokimyasal olarak tespit ve analizleri için öncelikle anti-miRNA molekülleri elektrot yüzeyine tutturulup ardından miRNA molekülleri ile etkileştirilerek hibridizasyona bağlı sinyal değişiklikleri incelenmektedir. miRNA molekülerinin tespitindeki hassaslıkta immobilizasyon süresi ve sıcaklığının yanı sıra yüzeye tutturulan anti-miRNA konsantrasyonuda kritik öneme sahiptir. Bu kapsamda yapılan bu çalışmada, sentetik olarak üretilen anti-miRNA (anti-miR451(G)) molekülleri farklı konsantrasyonlarda fosfat tampon çözeltisi kullanılarak seyreltilmiştir. Daha sonar bu çözeltiler yüzey-baskı elektrotlar (SPEs) yüzeylerine immobilize edilerek diferansiyel puls voltametri (DPV) metodu ile guanine oksidasyon sinyaleri incelenmiştir. Uygun konsantrasyonda hazırlanan çözelti damlatılmış SPE yüzeyler -18, +5 ve +25 oC sıcaklıklarda 1, 3, 14 ve 21 gün aralıklarda bekletilip DPV ölçümleri yapılarak optimum immobilizasyon süre ve sıcaklığı tespit edilmeye çalışılmıştır. DPV ölçüm sonuçlarına göre anti-miRNA’lardaki guanine oksidasyon sinyali immobilizasyon çözeltisinde bulunan anti-miRNA miktarına bağlı olarak belirli bir konsantrasyona kadar dramatik bir şekilde artmıştır daha sonra konsantrasyon artsa bile yüzeyin belirli bir doygunluk değerine ulaştığı için daha az artma eğilimine girmiştir. Ölçüm sonuçları değerlendirildiğinde SPE yüzeylerine tutturulmuş anti-miRNA moleküllerinin en yi bekleme sıcaklığının +5 oC olduğu gözlemlenmiştir.

Anahtar Kelimeler

miRNA detection, screen printed electrode, DPV measurement, guanine oxidation

Proje Numarası

118M231

Kaynakça

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