Nanoparticle-Based Proximity Ligation Assay for Ultrasensitive, Quantitative Detection of Protein Biomarkers

Hui Chen; Mary Crum; Dimple Chavan; Binh Vu; Katerina Kourentzi; Richard C. Willson

doi:10.1021/acsami.8b01377

Nanoparticle-Based Proximity Ligation Assay for Ultrasensitive, Quantitative Detection of Protein Biomarkers

Hui Chen, Mary Crum, Dimple Chavan, Binh Vu, Katerina Kourentzi, Richard C. Willson

Houston Methodist

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

Abstract

Proximity ligation assay (PLA) achieves extremely low limits of detection but requires the synthesis of antibody-DNA conjugates recognizing multiple target epitopes with appropriate proximity. In this work, we introduce a more generally applicable method by replacing antibody-DNA conjugates with nanoparticles which create ultradetectable PCR templates by capturing biotinylated oligonucleotides and catalyzing ligation. A competitive PCR readout was used to make the assay quantitative. We have demonstrated that NP-PLA can detect and quantitate human chorionic gonadotropin (hCG) levels as low as 2.6 fM (∼0.1 pg/mL), nearly 1000 times more sensitive than the current state of the art ELISA.

Original language	English (US)
Pages (from-to)	31845-31849
Number of pages	5
Journal	ACS Applied Materials and Interfaces
Volume	10
Issue number	38
DOIs	https://doi.org/10.1021/acsami.8b01377
State	Published - Sep 26 2018

Keywords

competitive polymerase chain reaction
immunoassay
nanoparticle
protein detection
proximity ligation assay

ASJC Scopus subject areas

General Materials Science

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10.1021/acsami.8b01377

Cite this

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abstract = "Proximity ligation assay (PLA) achieves extremely low limits of detection but requires the synthesis of antibody-DNA conjugates recognizing multiple target epitopes with appropriate proximity. In this work, we introduce a more generally applicable method by replacing antibody-DNA conjugates with nanoparticles which create ultradetectable PCR templates by capturing biotinylated oligonucleotides and catalyzing ligation. A competitive PCR readout was used to make the assay quantitative. We have demonstrated that NP-PLA can detect and quantitate human chorionic gonadotropin (hCG) levels as low as 2.6 fM (∼0.1 pg/mL), nearly 1000 times more sensitive than the current state of the art ELISA.",

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AU - Crum, Mary

AU - Chavan, Dimple

AU - Vu, Binh

AU - Kourentzi, Katerina

AU - Willson, Richard C.

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KW - competitive polymerase chain reaction

KW - immunoassay

KW - nanoparticle

KW - protein detection

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Nanoparticle-Based Proximity Ligation Assay for Ultrasensitive, Quantitative Detection of Protein Biomarkers

Abstract

Keywords

ASJC Scopus subject areas

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