Abstract
MRX34 has recently entered the clinic as the first therapeutic product based on a microRNA (miRNA) mimic. In order to measure drug concentrations in vivo, a quantitation method is needed that exhibits high precision, accuracy, and robustness. While most clinical applications for oligonucleotide therapeutics involve methods based on hybridization assays and liquid chromatography-tandem mass spectrometry, quantitative PCR (qPCR) is a less well-described approach. Here, we present an RT, qPCR, and analysis method to determine the tissue biodistribution of endogenous as well as a therapeutic, exogenous miRNA mimic therapeutic. Assay performance is demonstrated on multiple tissues from nonhuman primates dosed with MRX34.
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Abbreviations
- API:
-
Active pharmaceutical ingredient
- AVG:
-
Average
- cDNA:
-
Complementary DNA
- Ct:
-
Cycle threshold
- DNA:
-
Deoxyribonucleic acid
- dNTPs:
-
Deoxynucleotide triphosphates
- LLOQ:
-
Lower limit of quantitation
- miRNA:
-
microRNA
- NTC:
-
No-template control
- qRT-PCR:
-
Quantitative reverse transcription-polymerase chain reaction
- RNA:
-
Ribonucleic acid
- RT:
-
Reverse transcriptase
- tRNA:
-
transferRNA
- ULOQ:
-
Upper limit of quantitation
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© 2015 Springer Science+Business Media New York
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Kelnar, K., Bader, A.G. (2015). A qRT-PCR Method for Determining the Biodistribution Profile of a miR-34a Mimic. In: Walther, W., Stein, U. (eds) Gene Therapy of Solid Cancers. Methods in Molecular Biology, vol 1317. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2727-2_8
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DOI: https://doi.org/10.1007/978-1-4939-2727-2_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2726-5
Online ISBN: 978-1-4939-2727-2
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