I. Kira Astakhova

University of Southern Denmark, Denmark

Title: LNA/DNA probes for fluorescence sensing of nucleic acids and autoimmune antibodies


Kira Astakhova has completed her Ph.D. at the age of 23 years from Institute of bioorganic chemistry of Moscow, Russia, and postdoctoral studies from the Nucleic Acid Center, University of Southern Denmark. She is currently an Associate Professor at the Nucleic Acid Center, Denmark, a premier research institution focusing on synthetic nucleic acids. She has published more than 25 papers in reputed journals and has been serving as an editorial board member of repute. Her main research interests include nucleic acid chemistry and synthetic biology (LNA, DNA and RNA), nanobioscience and diagnostics of human diseases.


Several autoimmune disorders are characterized by production of antibodies against single- and double-stranded DNA. If not diagnosed and treated early, the autoimmune conditions can lead to serious health deterioration and even mortality. Th e sequence-specifi c autoimmune antibodies (autoantibodies) against single-stranded DNA have been thoroughly studied. In turn, non-sequence-specifi c autoantibodies against double-stranded DNA, a hallmark of autoimmune conditions such as antiphospholipide syndrome and systemic lupus erythematosus, have not been studied in detail. Generally, monitoring interactions of nucleic acids by fl uorescence is a convenient method in modern bioanalysis and can be performed under native conditions without additional equipment or procedures. Currently, fl uorescent oligonucleotides containing bright cyanine and xanthene dyes are oft en applied in bioanalysis of nucleic acids and proteins, including antibodies. Furthermore, affi nityenhancing locked nucleic acids containing 2-amino-LNA monomers with fl uorescent polyaromatic hydrocarbons (PAHs) attached at the 2-amino group provide high target binding affi nity and selectivity, remarkable fl uorescence quantum yields and brightness values. Another appealing aspect of LNA/DNA probes is their high potential as aptamers in selective binding of diverse proteins. Herein I will describe novel fl uorescent oligonucleotides for homogeneous (all-in-solution) detection of nucleic acids and autoimmune antibodies. Th e probes are prepared by effi cient click chemistry between novel alkyne-modifi ed locked nucleic acid (LNA) strands and a series of fl uorescent azides. Th e multiply labeled fl uorescent LNA/DNA probes prepared herein generally display high binding affi nity to complementary DNA/RNA, high quantum yields and, hence, high fl uorescence brightness values. With the novel fl uorescent probes, specifi c sensing of the monoclonal human autoantibody is performed. It makes the novel "clickable" LNA/DNA probes a very promising tool in molecular diagnostics and in studies of nucleic acids and autoantibodies against DNA. Th e latter are produced under autoimmune conditions including antiphospholipid syndrome and systemic lupus erythematosus.

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