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Barcode signal amplifying strategy for sensitive and accurate protein detection on LC-MS/MS
- Source :
- The Analyst. 146(5)
- Publication Year :
- 2021
-
Abstract
- Protein drugs showing strong pharmaceutical activity, high specificity, and low toxicity and side effects have drawn extensive attention in the field of life sciences and medicine. Precise evaluation of the function of these drugs requires accurate and sensitive detection methods. Here, we report a novel chromatography-tandem mass spectrometry (LC-MS/MS) method for sensitive and selective detection of protein drugs. Magnetic nanoparticles (Apt29@MNPs) were functionalized by thrombin aptamers, and quantum dots (Apt15@ss@QDs) were dual-functionalized with quantitative thrombin aptamers and small molecules with high ionization efficiency as the mass barcode. After Apt29@MNPs specifically purify and enrich thrombin from biological samples, they can form a nano "sandwich structure" when Apt15@ss@QDs are added, resulting in the release of the mass barcode for LC-MS/MS analysis via the cutting of the disulfide bond. Since there is a higher quantitative molecular ratio of mass barcode to thrombin in the nano-"sandwich structure", quantitative detection of thrombin with high sensitivity and selectivity can be achieved via the LC-MS/MS detection of the mass barcode with high ionization efficiency rather than thrombin, which effectively avoids the disadvantages of direct protein detection by mass spectrometry. The established method for thrombin detection shows a good linear relationship in a concentration range of 0.00115-1.15 nM with a limit of detection (LOD) of 0.0007 nM. The present work provides a new approach for the effective and sensitive quantitative analysis of protein drugs and would be of great significance in promoting the development of protein drugs and clinical applications.
- Subjects :
- Detection limit
Chromatography
Chemistry
Aptamer
Thrombin
Proteins
Barcode
Mass spectrometry
Biochemistry
Signal
Small molecule
Analytical Chemistry
law.invention
law
Limit of Detection
Tandem Mass Spectrometry
Electrochemistry
medicine
Environmental Chemistry
Quantitative analysis (chemistry)
Spectroscopy
medicine.drug
Chromatography, Liquid
Subjects
Details
- ISSN :
- 13645528
- Volume :
- 146
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- The Analyst
- Accession number :
- edsair.doi.dedup.....ecac46fe0a1de2c4ad38e46aa21ed268