Multiplexed Immunoassays for Proteins Using Magnetic Luminescent Nanoparticles for Internal Calibration

Details

• Personal Author:
  Dosev D ; Gee SJ ; Hammock BD ; Kennedy IM ; Nichkova M
• Description:
  Suspension arrays present a promising tool for multiplexed assays in large-scale screening applications. A simple and robust platform for quantitative multiprotein immunoanalysis has been developed with the use of magnetic Co:Nd:Fe(2)O(3)/luminescent Eu:Gd(2)O(3) core/shell nanoparticles (MLNPs) as a carrier. The magnetic properties of the MLNPs allow their manipulation by an external magnetic field in the separation and washing steps in the immunoassay. Their optical properties enable the internal calibration of the detection system. The multiplexed sandwich immunoassay involves dual binding events on the surface of the MLNPs functionalized with the capture antibodies. Secondary antibodies labeled with conventional organic dyes (Alexa Fluor) are used as reporters. The amount of the bound secondary antibody is directly proportional to the concentration of the analyte in the sample. In our approach, the fluorescence intensity of the reporter dye is related to the luminescence signal of the MLNPs. In this way, the intrinsic luminescence of the MLNPs serves as an internal standard in the quantitative immunoassay. The concept is demonstrated for a simultaneous immunoassay for three model proteins (human, rabbit, and mouse IgGs). The method uses a standard bench plate reader. It can be applied to disease diagnostics and to the detection of biological threats. [Description provided by NIOSH]
• Subjects:
  Biochemistry Chemistry Immune System Laboratories Occupational Health Safety
• Keywords:
  Antibody Response Biological Weapons Chemical Analysis Chemical Binding Immunology Laboratory Testing Magnetic Properties Nanotechnology Quantitative Analysis
• ISSN:
  0003-2697
• Document Type:
  Text
• Funding:
  Cooperative Agreement
• Genre:
  Journal Article
• Place as Subject:
  California ; OSHA Region 9
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  34-40
• Volume:
  369
• Issue:
  1
• NIOSHTIC Number:
  nn:20032996
• Citation:
  Anal Biochem 2007 Oct; 369(1):34-40
• Contact Point Address:
  Mikaela Nichkova, Department of Entomology and the UC Davis Cancer Center, One Shields Avenue, Davis, CA 95616
• Email:
  mnichkova@ucdavis.edu
• Federal Fiscal Year:
  2008
• NORA Priority Area:
  Agriculture, Forestry and Fishing
• Performing Organization:
  University of California - Davis
• Peer Reviewed:
  True
• Start Date:
  20010930
• Source Full Name:
  Analytical Biochemistry
• End Date:
  20270929
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:457c420782ce6f885c2c24424c19832efc5123aa96fc3eb1256659210d554223028ce67f7f72384a7afb4b301c127988e5aaae791366c55d49a534ccf2b53b3a
• Download URL:
  https://stacks.cdc.gov/view/cdc/188736/cdc_188736_DS1.pdf
• File Type:
  [PDF - 672.49 KB ]