Multiplex
biomolecular analysis with inductively coupled plasma
mass spectrometry (ICP-MS) becomes increasingly important in clinical
diagnosis and single cell analysis. However, the sensitivity of ICP-MS-based
immunoassay is only comparable or lower than those of fluorescence
methods at the present stage. Therefore, designing elemental tags
with a large number of metal atoms is necessary to achieve high-sensitive
detection. In this work, we proposed a new strategy to build up elemental
tag loading with hundreds of rare earth ions by coupling alkyne-DNA
chains with rare earth element (REE)-DOTA complexes a click chemistry
reaction. There are about 2 orders of magnitude improvement in sensitivity
compared with single metal-ion tags. DNA chains with multialkynyl
groups were facilely prepared by PCR synthesis. Moreover, the DNA-based
elemental tags own excellent water-solubility and biocompatibility.
The tags would be potentially applied to mass cytometry and clinical
diagnosis.
A combinatorial immunoassay method for biomarker detection based on a stable isotope tagging strategy was proposed. A multiplex immunoassay of 12 proteins could be achieved simultaneously and a combinatorial immunoassay was explored, which would be expected to satisfy the requirements of personalized detection.
The tool box of site-specific cleavage
for nucleic acid has been
an increasingly attractive subject. Especially, the recent emergence
of the orthogonally activatable DNA device is closely related to the
site-specific scission. However, most of these cleavage strategies
are based on exogenous assistance, such as laser irradiation. Endogenous
strategies are highly desirable for the orthogonally regulatable DNA
machine to explore the crucial intracellular biological process and
cell signal network. Here, we found that the accurate site-specific
cleavage reaction of phosphorothioate (PT) modified DNA by using myeloperoxidase
(MPO). A scissors-like mechanism by which MPO breaks PT modification
through chloride oxidation has been revealed. Furthermore, we have
successfully applied the scissors to activate PT-modified hairpin-DNA
machines to produce horseradish peroxidase (HRP)-mimicking DNAzyme
or initiate hybridization chain reaction (HCR) amplification. Since
MPO plays an important role in the pathway related to oxidative stress
in cells, through the HCR amplification activated by this tool box,
the oxidative stress in living cells has been robustly imaged. This
work proposes an accurate and endogenous site-specific cleavage tool
for the research of biostimuli and the construction of DNA molecular
devices.
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