TY  - GEN
A1  - Choi, Youngeun
A1  - Schmidt, Carsten
A1  - Tinnefeld, Philip
A1  - Bald, Ilko
A1  - Rödiger, Stefan
T1  - A new reporter design based on DNA origami nanostructures
for quantification of short oligonucleotides using microbeads
T2  - Postprints der Universität Potsdam : Mathematisch-naturwissenschaftliche Reihe
N2  - The DNA origami technique has great potential for the development of brighter and more sensitive reporters for fluorescence based detection schemes such as a microbead-based assay in diagnostic applications. The nanostructures can be programmed to include multiple dye molecules to enhance the measured signal as well as multiple probe strands to increase the binding strength of the target oligonucleotide to these nanostructures. Here we present a proof-of-concept study to quantify short oligonucleotides by developing a novel DNA origami based reporter system, combined with planar microbead assays. Analysis of the assays using the VideoScan digital imaging platform showed DNA origami to be a more suitable reporter candidate for quantification of the target oligonucleotides at lower concentrations than a conventional reporter that consists of one dye molecule attached to a single stranded DNA. Efforts have been made to conduct multiplexed analysis of different targets as well as to enhance fluorescence signals obtained from the reporters. We therefore believe that the quantification of short oligonucleotides that exist in low copy numbers is achieved in a better way with the DNA origami nanostructures as reporters.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 705 
KW  - nucleic-acids
KW  - hybridization
KW  - microrna
KW  - flourescence
KW  - biomarkers
KW  - platform
KW  - particle
KW  - binding
KW  - array
KW  - gene
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-428271
SN  - 1866-8372
IS  - 705
ER  - 
TY  - GEN
A1  - Czarnecka, Malgorzata
A1  - Weichelt, Ulrike
A1  - Rödiger, Stefan
A1  - Hanack, Katja
T1  - Novel Anti Double-Stranded Nucleic Acids Full-Length Recombinant Camelid Heavy-Chain Antibody for the Detection of miRNA
T2  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe
N2  - The discovery that certain diseases have specific miRNA signatures which correspond to disease progression opens a new biomarker category. The detection of these small non-coding RNAs is performed routinely using body fluids or tissues with real-time PCR, next-generation sequencing, or amplification-based miRNA assays. Antibody-based detection systems allow an easy onset handling compared to PCR or sequencing and can be considered as alternative methods to support miRNA diagnostic in the future. In this study, we describe the generation of a camelid heavy-chain-only antibody specifically recognizing miRNAs to establish an antibody-based detection method. The generation of nucleic acid-specific binders is a challenge. We selected camelid binders via phage display, expressed them as VHH as well as full-length antibodies, and characterized the binding to several miRNAs from a signature specific for dilated cardiomyopathy. The described workflow can be used to create miRNA-specific binders and establish antibody-based detection methods to provide an additional way to analyze disease-specific miRNA signatures.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 1274 
KW  - antibody
KW  - camelid antibody
KW  - heavy-chain-only antibody
KW  - miRNA
KW  - nucleic acids
KW  - novel biomarkers
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-569142
SN  - 1866-8372
SP  - 1
EP  - 18
ER  -