Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 49, Pages 17255-17262Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c04182
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Funding
- Cabinet Office (CAO), Cross-ministerial Strategic Innovation Promotion Program (SIP), Intelligent Processing Infrastructure of Cyber and Physical Systems (funding agency: NEDO)
- Japan Society for the Promotion of Science (JSPS) [18H02002, 22H05049]
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Researchers have developed an MTX detection system using chemically synthesizable homogeneous oligonucleotides. Through microbead-assisted capillary electrophoresis, they successfully identified MSmt7 with a similar level of specificity to antiMTX antibodies. This oligonucleotide can be coupled with a peroxidase-like hemin-DNAzyme to construct a bifunctional MTX sensing method with a low detection limit.
High-dose methotrexate (MTX) therapy is used to treat a wide variety of cancers such as leukemia and lymphoma, while the resulting high blood concentration of MTX faces a risk of life-threatening side effects, so it is essential to monitor the concentration carefully. Currently, the MTX concentration is measured using antibody-based kits in a clinical setting; however, the heterogeneity and batch-to-batch variation of antibodies potentially compromise the detection limit. Here, we developed MTX detection systems with chemically synthesizable homogeneous oligonucleotides. Microbead-assisted capillary electrophoresis (MACE)-SELEX against MTX successfully identified MSmt7 with a similar level of specificity to antiMTX antibodies within three rounds. The 3 '-end of MSmt7 was coupled to a peroxidase-like hemin-DNAzyme to construct a bifunctional oligonucleotide for MTX sensing, where MTX in 50% human serum was detected with a limit of detection (LoD) of 118 nM. Furthermore, amplifying the DNAzyme region with rolling circle amplification significantly improved the sensitivity with an LoD of 290 pM. Presented oligonucleotide-based MTX detection systems will pave the way for antibody-independent MTX detection with reliability and less cost in the laboratory and the clinic.
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