Aptasensor for Quantification of Leptin Through PCR Amplification of Short DNA-Aptamers.
Francesca Romana CavalloKhalid B MirzaSara de MateoKonstantin NikolicJesus Rodriguez-ManzanoChristofer ToumazouPublished in: ACS sensors (2021)
Protein quantification is traditionally performed through enzyme-linked immunosorbent assay (ELISA), which involves long preparation times. To overcome this, new approaches use aptamers as an alternative to antibodies. In this paper, we present a new approach to quantify proteins with short DNA aptamers through polymerase chain reaction (PCR) resulting in shorter protocol times with comparatively improved limits of detection. The proposed method includes a novel way to quantify both the target protein and the corresponding short DNA-aptamers simultaneously, which also allows us to fully characterize the performance of aptasensors. Human leptin is used as a target protein to validate this technique, because it is considered an important biomarker for obesity-related studies. In our experiments, we achieved the lowest limit of detection of 100 pg/mL within less than 2 h, a limit affected by the dissociation constant of the leptin aptamer, which could be improved by selecting a more specific aptamer. Because of the simple and inexpensive approach, this technique can be employed for Lab-On-Chip implementations and for rapid "on-site" quantification of proteins.
Keyphrases
- nucleic acid
- label free
- loop mediated isothermal amplification
- circulating tumor
- sensitive detection
- real time pcr
- protein protein
- single molecule
- cell free
- high throughput
- gold nanoparticles
- endothelial cells
- amino acid
- type diabetes
- binding protein
- metabolic syndrome
- randomized controlled trial
- insulin resistance
- weight loss
- quantum dots
- high resolution
- high fat diet induced
- mass spectrometry
- physical activity