Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin

Dosi Dosev, Mikaela Nichkova, Zhi Ya Ma, Shirley J. Gee, Bruce D. Hammock, Ian M. Kennedy

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Fluorescence techniques rely on measurement of relative fluorescence units and require calibration to obtain reliable and comparable quantitative data. Fluorescent immunoassays are a very sensitive and convenient method of choice for rapid detection of biotoxins, such as ricin. Here we present the application of magnetic luminescent nanoparticles (MLNPs) with a magnetic core of Fe 3O4 and a fluorescent shell of Eu:Gd2O 3 as carriers for a nanobead-immunoassay for the detection of ricin with internal calibration. A sandwich immunoassay for ricin was performed on the surface of the MLNPs. The particles were functionalized with capture polyclonal antibodies. Anti-ricin antibodies labeled with Alexa Fluor dye were used as the detecting antibodies. After magnetic extraction, the amount of ricin bound to the particle surface was quantified and related to the fluorescence signal of the nanoparticles. In this new platform, the MLNPs have three main functions: (1) a probe for the specific extraction of the target analyte from the sample; (2) a carrier in the quantitative immunoassay with magnetic separation; and (3) an internal standard in the fluorescence measurement of the dye reporter. The MLNPs serve as an internal control for the total analysis including extraction and assay performance. This approach eliminates the experimental error inherent in particle extraction and measurement of absolute organic dye fluorescence intensities. All fluorescent measurements were performed in a microplate reader. The standard curve for ricin had a dynamic range from 20 ng/ml to 100 μg/ml with a detection limit of 5 ng/ml. The configuration that has been developed can be easily adapted to a high throughput miniaturized system.

Original languageEnglish (US)
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6865
DOIs
StatePublished - 2008
EventNanoscale Imaging, Sensing, and Actuation for Biomedical Applications V - San Jose, CA, United States
Duration: Jan 21 2008Jan 23 2008

Other

OtherNanoscale Imaging, Sensing, and Actuation for Biomedical Applications V
CountryUnited States
CitySan Jose, CA
Period1/21/081/23/08

Fingerprint

Fluorescence
Calibration
Nanoparticles
Antibodies
Dyes
Magnetic cores
Magnetic separation
Assays
Throughput

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Dosev, D., Nichkova, M., Ma, Z. Y., Gee, S. J., Hammock, B. D., & Kennedy, I. M. (2008). Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 6865). [68650H] https://doi.org/10.1117/12.764030

Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin. / Dosev, Dosi; Nichkova, Mikaela; Ma, Zhi Ya; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 6865 2008. 68650H.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Dosev, D, Nichkova, M, Ma, ZY, Gee, SJ, Hammock, BD & Kennedy, IM 2008, Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 6865, 68650H, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications V, San Jose, CA, United States, 1/21/08. https://doi.org/10.1117/12.764030
Dosev D, Nichkova M, Ma ZY, Gee SJ, Hammock BD, Kennedy IM. Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 6865. 2008. 68650H https://doi.org/10.1117/12.764030
Dosev, Dosi ; Nichkova, Mikaela ; Ma, Zhi Ya ; Gee, Shirley J. ; Hammock, Bruce D. ; Kennedy, Ian M. / Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 6865 2008.
@inproceedings{a2ad95a323b34cadb566e2da1543dd1a,
title = "Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin",
abstract = "Fluorescence techniques rely on measurement of relative fluorescence units and require calibration to obtain reliable and comparable quantitative data. Fluorescent immunoassays are a very sensitive and convenient method of choice for rapid detection of biotoxins, such as ricin. Here we present the application of magnetic luminescent nanoparticles (MLNPs) with a magnetic core of Fe 3O4 and a fluorescent shell of Eu:Gd2O 3 as carriers for a nanobead-immunoassay for the detection of ricin with internal calibration. A sandwich immunoassay for ricin was performed on the surface of the MLNPs. The particles were functionalized with capture polyclonal antibodies. Anti-ricin antibodies labeled with Alexa Fluor dye were used as the detecting antibodies. After magnetic extraction, the amount of ricin bound to the particle surface was quantified and related to the fluorescence signal of the nanoparticles. In this new platform, the MLNPs have three main functions: (1) a probe for the specific extraction of the target analyte from the sample; (2) a carrier in the quantitative immunoassay with magnetic separation; and (3) an internal standard in the fluorescence measurement of the dye reporter. The MLNPs serve as an internal control for the total analysis including extraction and assay performance. This approach eliminates the experimental error inherent in particle extraction and measurement of absolute organic dye fluorescence intensities. All fluorescent measurements were performed in a microplate reader. The standard curve for ricin had a dynamic range from 20 ng/ml to 100 μg/ml with a detection limit of 5 ng/ml. The configuration that has been developed can be easily adapted to a high throughput miniaturized system.",
author = "Dosi Dosev and Mikaela Nichkova and Ma, {Zhi Ya} and Gee, {Shirley J.} and Hammock, {Bruce D.} and Kennedy, {Ian M.}",
year = "2008",
doi = "10.1117/12.764030",
language = "English (US)",
isbn = "9780819470409",
volume = "6865",
booktitle = "Progress in Biomedical Optics and Imaging - Proceedings of SPIE",

}

TY - GEN

T1 - Magnetic luminescent nanoparticles as internal calibration for an immunoassay for ricin

AU - Dosev, Dosi

AU - Nichkova, Mikaela

AU - Ma, Zhi Ya

AU - Gee, Shirley J.

AU - Hammock, Bruce D.

AU - Kennedy, Ian M.

PY - 2008

Y1 - 2008

N2 - Fluorescence techniques rely on measurement of relative fluorescence units and require calibration to obtain reliable and comparable quantitative data. Fluorescent immunoassays are a very sensitive and convenient method of choice for rapid detection of biotoxins, such as ricin. Here we present the application of magnetic luminescent nanoparticles (MLNPs) with a magnetic core of Fe 3O4 and a fluorescent shell of Eu:Gd2O 3 as carriers for a nanobead-immunoassay for the detection of ricin with internal calibration. A sandwich immunoassay for ricin was performed on the surface of the MLNPs. The particles were functionalized with capture polyclonal antibodies. Anti-ricin antibodies labeled with Alexa Fluor dye were used as the detecting antibodies. After magnetic extraction, the amount of ricin bound to the particle surface was quantified and related to the fluorescence signal of the nanoparticles. In this new platform, the MLNPs have three main functions: (1) a probe for the specific extraction of the target analyte from the sample; (2) a carrier in the quantitative immunoassay with magnetic separation; and (3) an internal standard in the fluorescence measurement of the dye reporter. The MLNPs serve as an internal control for the total analysis including extraction and assay performance. This approach eliminates the experimental error inherent in particle extraction and measurement of absolute organic dye fluorescence intensities. All fluorescent measurements were performed in a microplate reader. The standard curve for ricin had a dynamic range from 20 ng/ml to 100 μg/ml with a detection limit of 5 ng/ml. The configuration that has been developed can be easily adapted to a high throughput miniaturized system.

AB - Fluorescence techniques rely on measurement of relative fluorescence units and require calibration to obtain reliable and comparable quantitative data. Fluorescent immunoassays are a very sensitive and convenient method of choice for rapid detection of biotoxins, such as ricin. Here we present the application of magnetic luminescent nanoparticles (MLNPs) with a magnetic core of Fe 3O4 and a fluorescent shell of Eu:Gd2O 3 as carriers for a nanobead-immunoassay for the detection of ricin with internal calibration. A sandwich immunoassay for ricin was performed on the surface of the MLNPs. The particles were functionalized with capture polyclonal antibodies. Anti-ricin antibodies labeled with Alexa Fluor dye were used as the detecting antibodies. After magnetic extraction, the amount of ricin bound to the particle surface was quantified and related to the fluorescence signal of the nanoparticles. In this new platform, the MLNPs have three main functions: (1) a probe for the specific extraction of the target analyte from the sample; (2) a carrier in the quantitative immunoassay with magnetic separation; and (3) an internal standard in the fluorescence measurement of the dye reporter. The MLNPs serve as an internal control for the total analysis including extraction and assay performance. This approach eliminates the experimental error inherent in particle extraction and measurement of absolute organic dye fluorescence intensities. All fluorescent measurements were performed in a microplate reader. The standard curve for ricin had a dynamic range from 20 ng/ml to 100 μg/ml with a detection limit of 5 ng/ml. The configuration that has been developed can be easily adapted to a high throughput miniaturized system.

UR - http://www.scopus.com/inward/record.url?scp=42149124320&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=42149124320&partnerID=8YFLogxK

U2 - 10.1117/12.764030

DO - 10.1117/12.764030

M3 - Conference contribution

SN - 9780819470409

VL - 6865

BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE

ER -