Tetraspanin immunoassay for the detection of extracellular vesicles and renal cell carcinoma




Khan Misba, Islam Md. Khirul, Rahman Mafiur, Dhondt Bert, Quintero Ileana, Puhka Maija, Jaakkola Panu M., Lamminmäki Urpo, Leivo Janne

PublisherWiley-VCH Verlag GmbH & Co. KGaA

2024

Nano select

NANO SELECT

2400018

5

9

2688-4011

DOIhttps://doi.org/10.1002/nano.202400018(external)

https://doi.org/10.1002/nano.202400018(external)

https://research.utu.fi/converis/portal/detail/Publication/393558373(external)



Half of patients with renal cell carcinoma (RCC) develop metastases. New and noninvasive biomarkers are needed for the diagnosis of RCC. The study aims to develop an EV-based assay for the detection of RCC using a highly sensitive nanoparticle-aided time-resolved fluorescence immunoassay (NP-TRFIA). To confirm the presence of tetraspanins on EVs, size exclusion chromatography is used to separate EV- and PE-fractions from RCC4, 786-O, and HEK293 cell lines. EV- and PE-fractions are quantified using NP-TRFIA assays established for CD9, CD63, CD81, and CD151. Tetraspanins are measured from RCC CCM and serum samples of RCC (n = 14), benign (n = 17), and healthy (n = 9) individuals. Among the tetraspanins, CD63 exhibits 3-5-fold higher expression on RCC4 and 786-O CCM compared to HEK293. A sandwich CD63-CD63 assay demonstrates significant discrimination of RCC patients from benign (p = 0.0003), and healthy (p = 0.005) individuals, respectively. Similarly, the CD81-CD81 assay also enables significant separation of RCC patients compared to benign (p = 0.014), and healthy (p = 0.003) controls, respectively. This suggests that RCC cell lines and serum of RCC patients show higher amounts of CD63- and CD81-EVs compared to controls. Detection of these EVs using NP-TRFIA approach may play a vital role in the detection of RCC.


Last updated on 2024-17-12 at 10:47