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Schmieder, Stefan; Weisspflog, Janek; Danz, Norbert; Hübner, Max; Kreth, Simone; Klotzbach, Udo; Sonntag, Frank (2017): Ultrasensitive SPR detection of miRNA-93 using antibody-enhanced and enzymatic signal amplification. In: Engineering in Life Sciences, Vol. 17, No. 12: pp. 1264-1270
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Abstract

MiRNAs are endogenous noncoding RNAmolecules. They play important gene-regulatory roles by binding to the mRNA of target genes thereby leading to either transcript degradation or translational repression. In virtually all diseases, distinct alterations of miRNA expression profiles have been found thus suggesting miRNAs as interesting biomarkers. Here, we present an SPR biosensor that utilizes disposable, injection-molded sensor chip/microfluidic hybrids combined with a lateral imaging optical system for parallel analysis of three one-dimensional spot arrays to detect miRNA-93. To increase the sensitivity of the biosensor we used two different amplification strategies. By adding an RNA-DNA-hybrid antibody for primary signal amplification, a limit of detection of 10 pmol/L was achieved. Based on that method we demonstrate the detection of miRNA-93 in total RNA lysate from HEK-293 cells. Utilizing an enzymatic signal amplification with Poly(A) polymerase, the sensitivity could be increased even further leading to a limit of detection of 1 fmol/L.