Document Detail


Novel nonseparation sandwich-type electrochemical enzyme immunoassay system for detecting marker proteins in undiluted blood.
MedLine Citation:
PMID:  7544708     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A novel nonseparation electrochemical enzyme immunoassay (NEEIA) for detecting marker proteins in undiluted blood is described. The approach is based on preferential electrochemical measurement of surface-bound enzyme-labeled reporter antibody (E-Ab), relative to an excess of this reagent in the sample solution. NEEIAs are carried out on microporous membranes coated with a thin, circular area of gold. The gold serves simultaneously as a working electrode and solid phase for immobilized capture anti-protein antibodies. In the assay, analyte protein is incubated concurrently with the Ab-coated gold surface and excess E-Ab conjugate. Detection of bound E-Ab is achieved by introducing the substrate for the enzyme through the back side of the membrane. The product of bound E-Ab is detected immediately by oxidation or reduction at the gold electrode, and the resulting current is proportional to the concentration of protein in the sample. The feasibility of the NEEIA approach is demonstrated via the detection of prostate-specific antigen in undiluted plasma samples (n = 64), with alkaline phosphatase as the label. Use of multiple gold films deposited on the same porous membrane to perform simultaneous NEEIAs is also described.
Authors:
M E Meyerhoff; C Duan; M Meusel
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Clinical chemistry     Volume:  41     ISSN:  0009-9147     ISO Abbreviation:  Clin. Chem.     Publication Date:  1995 Sep 
Date Detail:
Created Date:  1995-10-03     Completed Date:  1995-10-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9421549     Medline TA:  Clin Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1378-84     Citation Subset:  IM    
Affiliation:
Department of Chemistry, University of Michigan, Ann Arbor 48109, USA.
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal
Biological Markers / blood*
Blood Proteins / analysis*
Chorionic Gonadotropin / blood
Electrochemistry
Electrodes
Gold
Humans
Immunoenzyme Techniques*
Immunoglobulin G
Oxidation-Reduction
Prostate-Specific Antigen / blood
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Biological Markers; 0/Blood Proteins; 0/Chorionic Gonadotropin; 0/Immunoglobulin G; 7440-57-5/Gold; EC 3.4.21.77/Prostate-Specific Antigen

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