Simultaneous immunoassay of phosphorylated proteins based on apoferritin templated metallic phosphates as voltammetrically distinguishable signal reporters

Details

• Personal Author:
  Du D ; Ge X ; Lin Y ; Zhang A
• Description:
  A novel electrochemical immunosensor has been developed to detect phosphorylated proteins, phospho-p53(15) and phospho-p53(392), simultaneously. Different apoferritin templated metal phosphates were used as distinguishable signal reporters (apoferritin templated cadmium phosphates (ATCP) and apoferritin templated lead phosphates (ATLP)) to enhance the detection sensitivity. Here, magnetic Fe3O4 nanoparticles functionalized phospho-p53(15) capture antibody (MP-p53(15)c-Ab) and phospho-p53(392) capture antibody (MP-p53(392)c-Ab), respectively, were used to specifically capture phospho-p53(15) and phospho-p53(392) antigens, followed by immunorecognition with p53(15) detection antibody (p53(15)d-Ab) and p53(392) detection antibody (p53(392)d-Ab) to form sandwich-like immunocomplexes. SiO2@Au nanocomposites served as nanocarriers for co-immobilization of both d-Ab and signal reporters (ATCP/SiO2@Au/p53(15)d-Ab, ATLP/SiO2@Au/p53(392)d-Ab), which greatly amplified the detection signal. The distinguished current responses were achieved by electrochemical detection of cadmium ions and lead ions with square wave voltammetry (SWV) after dissolution with acid. The proposed immunoassay exhibited high sensitivity and selectivity for the detection of phospho-p53(15) and phospho-p53(392) simultaneously. The linear relationships between electrochemical responses and the concentrations of phospho-p53(15) and phospho-p53(392) were obtained in the range of 0.1-20 ng/mL and 0.05-20 ng/mL, respectively. The detection limits were 0.05 and 0.02 ng/mL (S/N=3), respectively. This strategy provides a new platform for simultaneous immunoassay of multiple protein biomarkers. [Description provided by NIOSH]
• Subjects:
  Antibodies Antigens Biomarkers Cadmium Ferrous Compounds Immune System Lead Metals Occupational Health Phosphoric Acids Safety

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• Keywords:
  Apoferritin Templated Metallic Phosphates Author Keywords: Simultaneous Immunoassay Biosensors Cadmium Compounds Cell Signaling Cellular Reactions Chemical Agent Detectors Detectors Electrochemical Analysis Electrochemical Detection Immune Reaction Immunoassays Immunochemistry Iron Lead Compounds Metal Compounds Metallic Ions Nanocomposites Nanoparticles Nanotechnology Phoaphorylated Protein Phosphates Proteins Sensitivity Testing Sensors Volumetric Analysis

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• ISSN:
  0956-5663
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 10 ; Washington
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  201-207
• Volume:
  80
• NIOSHTIC Number:
  nn:20049091
• Citation:
  Biosens Bioelectron 2016 Jun; 80:201-207
• Contact Point Address:
  Dan Du, Key Laboratory of Pesticide and Chemical Biology of Ministry of Education,College of Chemistry, Central China Normal University, Wuhan 430079, PR China
• Email:
  dan.du@mail.ccnu.edu.cn
• Federal Fiscal Year:
  2016
• NORA Priority Area:
  Agriculture, Forestry and Fishing
• Performing Organization:
  Washington State University
• Peer Reviewed:
  True
• Start Date:
  20150901
• Source Full Name:
  Biosensors and Bioelectronics
• End Date:
  20180831
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:f37e47ea73936984ba566e90e0b5a9992282981784a8ef7780e78120323dc36b99b6c25dbf36ebbc287773fa332b96795c20106ef6306c6c66000116c62d3dd7
• Download URL:
  https://stacks.cdc.gov/view/cdc/203664/cdc_203664_DS1.pdf
• File Type:
  [PDF - 1.85 MB ]