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| Content Provider | Royal Society of Chemistry (RSC) |
|---|---|
| Author | Wan, Debin Lian, Wenhui Yao, Wenbin Chen, Changbao Liu, Shuying Su, Rui Shi, Lei Xu, Niusheng Yang, Hongmei Wang, Yihan |
| Copyright Year | 2017 |
| Abstract | Conventional strategies for the screening of DNA triplex binders cannot be used for complicated samples, such as ligand libraries created by combinatorial chemistry or from natural product extracts. In the current study, an ultra-high-performance liquid chromatography coupled with an Orbitrap mass spectrometry (UHPLC-Orbitrap-MS)-based approach, which we call peak area-fading (PAF) UHPLC-Orbitrap-MS and was designed for just such a purpose, is reported. The triplex DNA modified 96-well plate and the single stranded oligonucleotide modified 96-well plate (as control) were incubated with ligand libraries, and the unbound ligands were directly determined via UHPLC-ESI-MS. The binders were detected through the decrease (fading) in the peak areas compared to those of the control group. Several factors, such as incubation time, incubation temperature, and buffer, which might affect the binding affinity and reproducibility, were optimized. The potential of the approach was examined using the extracts of Rhizoma Coptidis and Phellodendron chinense Schneid cortexe. The triplex DNA-binding capabilities of the five components (epiberberine, coptisine, jatrorrhizine, berberrubine, and columbamine) were found for the first time, indicating their efficiency for the analysis of complicated samples. In contrast to our previous study, which suffered from a serious drawback of poor reproducibility, this method is more robust and more suitable for high-throughput measurements, opening a new experimental strategy in assessing large libraries of potential drug candidates that work by forming a drug/DNA complex. |
| Starting Page | 670 |
| Ending Page | 675 |
| Page Count | 6 |
| File Format | HTM / HTML PDF |
| ISSN | 00032654 |
| Volume Number | 142 |
| Issue Number | 4 |
| Journal | The Analyst |
| DOI | 10.1039/c6an01974a |
| Language | English |
| Publisher | Royal Society of Chemistry |
| Access Restriction | Open |
| Subject Keyword | Phellodendron Ligand Combinatorial chemistry Natural product Pakistan Air Force Orbitrap High-performance liquid chromatography Coptisine PAF DNA Jatrorrhizine Oligonucleotide Mass spectrometry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry Electrochemistry |
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