Back to Search
Start Over
Click Chemistry Mediated Eu-Tagging: Activity-Based Specific Quantification and Simultaneous Activity Evaluation of CYP3A4 Using 153Eu Species-Unspecific Isotope Dilution Inductively Coupled Plasma Mass Spectrometry
- Source :
- Analytical Chemistry. 86:3688-3692
- Publication Year :
- 2014
- Publisher :
- American Chemical Society (ACS), 2014.
-
Abstract
- P450 3A4 (CYP3A4) is one of the most important isoforms in the human cytochrome P450 superfamily. It was used as an example in this proof-of-concept study in order to demonstrate an activity-based labeling and then click chemistry (CC) mediated element-tagging strategy for simultaneously specific quantification and activity measurement of an enzyme using species-unspecific isotope dilution inductively coupled plasma mass spectrometry (SUID ICPMS). A dual functional hexynylated 17α-ethynylestradiol activity-based probe was synthesized for specifically labeling CYP3A4 and then CC-mediated Eu-tagging with an azido-DOTA-Eu complex for CYP3A4 quantification and activity measurement in human liver microsome and serum samples using (153)Eu SUID ICPMS. The LOD (3σ) of CYP3A4 reached 20.3 fmol when monitoring (151/153)Eu ICPMS signals, in addition to the merits of specificity and simultaneous activity measurement achieved. We believe that this activity-based CC-mediated element-tagging strategy will liberate more potential advantages of ICPMS in bioanalysis.
- Subjects :
- Radioisotopes
Radioisotope Dilution Technique
Bioanalysis
Chromatography
Estradiol
Isotope
Chemistry
Isotope dilution
Analytical Chemistry
Label-free quantification
Europium
Isotopes
Steroid Hydroxylases
Click chemistry
Cytochrome P-450 CYP3A
Humans
Click Chemistry
Multiplex
Quantitative analysis (chemistry)
Inductively coupled plasma mass spectrometry
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 86
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....04cd59d335b0094c84d51e6c3ae203c5