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

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

17 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

Materials Science(all)

Access to Document

10.1021/acsami.8b01377

Cite this

@article{86498a0396ca49a5b04d6c3dcac763b4,

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

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.",

keywords = "competitive polymerase chain reaction, immunoassay, nanoparticle, protein detection, proximity ligation assay",

author = "Hui Chen and Mary Crum and Dimple Chavan and Binh Vu and Katerina Kourentzi and Willson, {Richard C.}",

note = "Funding Information: *E-mail: willson@uh.edu. ORCID Richard C. Willson: 0000-0002-0900-6148 Funding We gratefully acknowledge support from National Institute of Allergy and Infectious Diseases, National Institutes of Health, http://www.niaid.nih.gov (Grant 1R21AI111120-01A1);National Science Foundation, www.nsf.gov (Grant CBET-1511789); and Cancer Prevention & Research Institute of Texas, www.cprit.state.tx.us (Grant RP150343). Notes The authors declare the following competing financial interest(s): Several of the authors of this manuscript are named inventors on pending IP which overlaps the topics of the manuscript. Publisher Copyright: {\textcopyright} 2018 American Chemical Society.",

year = "2018",

month = sep,

day = "26",

doi = "10.1021/acsami.8b01377",

language = "English (US)",

volume = "10",

pages = "31845--31849",

journal = "ACS Applied Materials and Interfaces",

issn = "1944-8244",

publisher = "American Chemical Society",

number = "38",

}

TY - JOUR

T1 - Nanoparticle-Based Proximity Ligation Assay for Ultrasensitive, Quantitative Detection of Protein Biomarkers

AU - Chen, Hui

AU - Crum, Mary

AU - Chavan, Dimple

AU - Vu, Binh

AU - Kourentzi, Katerina

AU - Willson, Richard C.

N1 - Funding Information: *E-mail: willson@uh.edu. ORCID Richard C. Willson: 0000-0002-0900-6148 Funding We gratefully acknowledge support from National Institute of Allergy and Infectious Diseases, National Institutes of Health, http://www.niaid.nih.gov (Grant 1R21AI111120-01A1);National Science Foundation, www.nsf.gov (Grant CBET-1511789); and Cancer Prevention & Research Institute of Texas, www.cprit.state.tx.us (Grant RP150343). Notes The authors declare the following competing financial interest(s): Several of the authors of this manuscript are named inventors on pending IP which overlaps the topics of the manuscript. Publisher Copyright: © 2018 American Chemical Society.

PY - 2018/9/26

Y1 - 2018/9/26

N2 - 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.

AB - 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.

KW - competitive polymerase chain reaction

KW - immunoassay

KW - nanoparticle

KW - protein detection

KW - proximity ligation assay

UR - http://www.scopus.com/inward/record.url?scp=85053640431&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85053640431&partnerID=8YFLogxK

U2 - 10.1021/acsami.8b01377

DO - 10.1021/acsami.8b01377

M3 - Article

C2 - 30168312

AN - SCOPUS:85053640431

VL - 10

SP - 31845

EP - 31849

JO - ACS Applied Materials and Interfaces

JF - ACS Applied Materials and Interfaces

SN - 1944-8244

IS - 38

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this