Journal
BIOSENSORS & BIOELECTRONICS
Volume 141, Issue -, Pages -Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2019.111403
Keywords
Fluorescence resonance energy transfer biosensor; Peptide modified upconversion nanoparticle; Caspase-9 activity; Upconversion luminescence imaging
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Funding
- National Natural Science Foundation of China [21775145]
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In the present work, peptide-functionalized upconversion@silica nanoparticle (termed as UCNP@SiO2@Cy5-pep)-based fluorescence resonance energy transfer (FRET) sensing platform has been fabricated for the detection of caspase-9 activity in vitro and in vivo. A Cy5 labeled peptide containing specific motif LEHD for caspase-9 cleavage was designed and conjugated with UCNP@SiO2 through covalent attachment. The red upconversion luminescence (UCL) emission of UCNP@SiO2 can be quenched by Cy5 while the green UCL emission of UCNP@SiO2 remains undisturbed. After the cleavage of LEHD by caspase-9, the Cy5 departed from UCNP@SiO2 surface, resulting in recovery of red UCL emission of UCNP@SiO2. The UCNP@SiO2@Cy5-pep has been successfully used to monitoring the changes of caspase-9 activity levels in apoptotic cancerous cells (MG-63 and SW480) by cisplatin-induction. Under same experimental conditions, it is found that the intracellular caspase-9 activity level of cisplatin-treated MG-63 cells is higher than that of cisplatin-treated SW480. This result is consistent with that of commercial caspase-9 activity kits. The UCL signal intensity ratio of red emission to green emission of UCNP (termed as R/G) is linearly dependent on the amount of MG-63 cells within the range of 5 x 10(3) to 1 x 10(6) cells (i.e., 0.5-100 U mL(-1) caspase-9) with a limit of detection (LOD) of 675 cells (i.e., 0.068 U mL(-1) caspase-9). Furthermore, the practicability of UCNP@SiO2@Cy5-pep is demonstrated by detection of caspase-9 activities in tumor tissues in vivo, and satisfactory results are obtained.
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