Neutral DNA-avidin nanoparticles as ultrasensitive reporters in immuno-PCR

Dimple Chavan; Hui Chen; Mary Crum; Binh Vu; Mohammad Safari; Maxwell Smith; Peter Vekilov; Jacinta C. Conrad; Katerina Kourentzi; Richard C. Willson

doi:10.1039/d0an00134a

Neutral DNA-avidin nanoparticles as ultrasensitive reporters in immuno-PCR

Dimple Chavan, Hui Chen, Mary Crum, Binh Vu, Mohammad Safari, Maxwell Smith, Peter Vekilov, Jacinta C. Conrad, Katerina Kourentzi, Richard C. Willson

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

1 Scopus citations

Abstract

We have developed an immuno-PCR based diagnostic platform which couples detection antibodies to self-assembled, ultra-detectable DNA-avidin nanoparticles stabilized with poly(ethylene glycol) to link DNA amplification to target protein concentration. Electrostatic neutralization and cloaking of the PCR-amplifiable DNA labels by avidin and PEG coating reduces non-specific "stickiness" and enhances assay sensitivity. We further optimized the detectability of the nanoparticles by incorporating four repeats of a unique synthetic DNA PCR target into each nanoparticle. Using human chorionic gonadotropin hormone (hCG) as a model analyte, this platform was able to quantitate the target hCG protein in femtomolar concentrations using only standard laboratory equipment.

Original language	English (US)
Pages (from-to)	4942-4949
Number of pages	8
Journal	Analyst
Volume	145
Issue number	14
DOIs	https://doi.org/10.1039/d0an00134a
State	Published - Jul 21 2020

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Environmental Chemistry
Spectroscopy
Electrochemistry

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title = "Neutral DNA-avidin nanoparticles as ultrasensitive reporters in immuno-PCR",

abstract = "We have developed an immuno-PCR based diagnostic platform which couples detection antibodies to self-assembled, ultra-detectable DNA-avidin nanoparticles stabilized with poly(ethylene glycol) to link DNA amplification to target protein concentration. Electrostatic neutralization and cloaking of the PCR-amplifiable DNA labels by avidin and PEG coating reduces non-specific {"}stickiness{"} and enhances assay sensitivity. We further optimized the detectability of the nanoparticles by incorporating four repeats of a unique synthetic DNA PCR target into each nanoparticle. Using human chorionic gonadotropin hormone (hCG) as a model analyte, this platform was able to quantitate the target hCG protein in femtomolar concentrations using only standard laboratory equipment.",

author = "Dimple Chavan and Hui Chen and Mary Crum and Binh Vu and Mohammad Safari and Maxwell Smith and Peter Vekilov and Conrad, {Jacinta C.} and Katerina Kourentzi and Willson, {Richard C.}",

note = "Funding Information: We gratefully acknowledge support from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, http://www.niaid.nih.gov (Grant 1R21AI111120-01A1); National Science Foundation, http://www.nsf.gov (Grant CBET-1511789); and Cancer Prevention & Research Institute of Texas, http://www.cprit.state.tx.us (Grant RP150343). Publisher Copyright: {\textcopyright} 2020 The Royal Society of Chemistry.",

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doi = "10.1039/d0an00134a",

language = "English (US)",

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AU - Chavan, Dimple

AU - Chen, Hui

AU - Crum, Mary

AU - Vu, Binh

AU - Safari, Mohammad

AU - Smith, Maxwell

AU - Vekilov, Peter

AU - Conrad, Jacinta C.

AU - Kourentzi, Katerina

AU - Willson, Richard C.

N1 - Funding Information: We gratefully acknowledge support from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, http://www.niaid.nih.gov (Grant 1R21AI111120-01A1); National Science Foundation, http://www.nsf.gov (Grant CBET-1511789); and Cancer Prevention & Research Institute of Texas, http://www.cprit.state.tx.us (Grant RP150343). Publisher Copyright: © 2020 The Royal Society of Chemistry.

PY - 2020/7/21

Y1 - 2020/7/21

N2 - We have developed an immuno-PCR based diagnostic platform which couples detection antibodies to self-assembled, ultra-detectable DNA-avidin nanoparticles stabilized with poly(ethylene glycol) to link DNA amplification to target protein concentration. Electrostatic neutralization and cloaking of the PCR-amplifiable DNA labels by avidin and PEG coating reduces non-specific "stickiness" and enhances assay sensitivity. We further optimized the detectability of the nanoparticles by incorporating four repeats of a unique synthetic DNA PCR target into each nanoparticle. Using human chorionic gonadotropin hormone (hCG) as a model analyte, this platform was able to quantitate the target hCG protein in femtomolar concentrations using only standard laboratory equipment.

AB - We have developed an immuno-PCR based diagnostic platform which couples detection antibodies to self-assembled, ultra-detectable DNA-avidin nanoparticles stabilized with poly(ethylene glycol) to link DNA amplification to target protein concentration. Electrostatic neutralization and cloaking of the PCR-amplifiable DNA labels by avidin and PEG coating reduces non-specific "stickiness" and enhances assay sensitivity. We further optimized the detectability of the nanoparticles by incorporating four repeats of a unique synthetic DNA PCR target into each nanoparticle. Using human chorionic gonadotropin hormone (hCG) as a model analyte, this platform was able to quantitate the target hCG protein in femtomolar concentrations using only standard laboratory equipment.

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Neutral DNA-avidin nanoparticles as ultrasensitive reporters in immuno-PCR

Abstract

ASJC Scopus subject areas

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