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Ultrasensitive quantification of PD-L1+ extracellular vesicles in melanoma patient plasma using a parallelized high throughput droplet digital assay.
Shen, Hanfei; Atiyas, Yasemin; Yang, Zijian; Lin, Andrew A; Yang, Jingbo; Liu, Diao; Park, Juhwan; Guo, Wei; Issadore, David A.
Afiliación
  • Shen H; Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA. issadore@seas.upenn.edu.
  • Atiyas Y; Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA. issadore@seas.upenn.edu.
  • Yang Z; Department of Radiology, School of Medicine, Stanford University, Stanford, California, USA.
  • Lin AA; Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA. issadore@seas.upenn.edu.
  • Yang J; Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA, USA.
  • Liu D; Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA, USA.
  • Park J; Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA. issadore@seas.upenn.edu.
  • Guo W; Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA, USA.
  • Issadore DA; Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA. issadore@seas.upenn.edu.
Lab Chip ; 24(14): 3403-3411, 2024 07 10.
Article en En | MEDLINE | ID: mdl-38899443
ABSTRACT
The expression of programmed death-ligand 1 (PD-L1) on extracellular vesicles (EVs) is an emerging biomarker for cancer, and has gained particular interest for its role mediating immunotherapy. However, precise quantification of PD-L1+ EVs in clinical samples remains challenging due to their sparse concentration and the enormity of the number of background EVs in human plasma, limiting applicability of conventional approaches. In this study, we develop a high-throughput droplet-based extracellular vesicle analysis (DEVA) assay for ultrasensitive quantification of EVs in plasma that are dual positive for both PD-L1 and tetraspanin (CD81) known to be expressed on EVs. We achieve a performance that significantly surpasses conventional approaches, demonstrating 360× enhancement in the limit of detection (LOD) and a 750× improvement in the limit of quantitation (LOQ) compared to conventional plate enzyme-linked immunoassay (ELISA). Underlying this performance is DEVA's high throughput analysis of individual EVs one at a time and the high specificity to targeted EVs versus background. We achieve a 0.006% false positive rate per droplet by leveraging avidity effects that arise from EVs having multiple copies of their target ligands on their surface. We use parallelized optofluidics to rapidly process 10 million droplets per minute, ∼100× greater than conventional approaches. A validation study on a cohort of 14 patients with melanoma confirms DEVA's ability to match conventional ELISA measurements with reduced plasma sample volume and without the need for prior EV purification. This proof-of-concept study demonstrates DEVA's potential for clinical utility to enhance prognosis as well as guide treatment for cancer.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Antígeno B7-H1 / Vesículas Extracelulares / Melanoma Límite: Humans Idioma: En Revista: Lab Chip Asunto de la revista: BIOTECNOLOGIA / QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Antígeno B7-H1 / Vesículas Extracelulares / Melanoma Límite: Humans Idioma: En Revista: Lab Chip Asunto de la revista: BIOTECNOLOGIA / QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido