Heterogeneous Enzyme Immunoassay with Amperometric Detection

Author(s):  
Kenneth R. Wehmeyer ◽  
Matthew J. Doyle ◽  
H. Brian Halsall ◽  
William R. Heineman
1990 ◽  
Vol 36 (11) ◽  
pp. 1941-1944 ◽  
Author(s):  
Y Xu ◽  
B Halsall ◽  
W R Heineman

Abstract A sandwich-type heterogeneous enzyme immunoassay with flow-injection analysis for alpha-fetoprotein (AFP) in human serum has been developed. 4-Aminophenol, the product of enzymatic reaction, is detected amperometrically. The interassay CV for this electrochemical enzyme immunoassay was less than 8.2%, with a minimum detection limit for AFP of 0.163 micrograms/L. The calibration curve had a linear range of 0.316-100 micrograms/L. Studies with 48 human maternal serum samples, comparing results by this method with those by a commercial kit, showed a good correlation (r = 0.961). This procedure provides an alternative method for determining low concentrations of AFP in human maternal serum.


2001 ◽  
Vol 120 (5) ◽  
pp. A492-A493 ◽  
Author(s):  
E HAINDL ◽  
H BENESCH ◽  
A FINCK ◽  
V MUEHISTEIN ◽  
A LEODOLTER ◽  
...  

2001 ◽  
Vol 120 (5) ◽  
pp. A492-A492
Author(s):  
Y YEE ◽  
E YIP ◽  
T QUE ◽  
K LI ◽  
C LEE ◽  
...  

1988 ◽  
Vol 59 (02) ◽  
pp. 310-315 ◽  
Author(s):  
P W Koppert ◽  
E Hoegee-de Nobel ◽  
W Nieuwenhuizen

SummaryWe have developed a sandwich-type enzyme immunoassay (EIA) for the quantitation of fibrin degradation products (FbDP) in plasma with a time-to-result of only 45 minutes.* The assay is based on the combination of the specificities of two monoclonal antibodies (FDP-14 and DD-13), developed in our institute. FDP-14, the capture antibody, binds both fibrinogen degradation products (FbgDP) and FbDP, but does not react with the parent fibrin(ogen) molecules. It has its epitope in the E-domain of the fibrinogen molecule on the Bβ-chain between amino acids 54-118. Antibody DD-13 was raised using D-dimer as antigen and is used as a tagging antibody, conjugated with horse-radish peroxidase. A strong positive reaction is obtained with a whole blood clot lysate (lysis induced by tissue-type plasminogen activator) which is used as a standard. The EIA does virtually not detect FbgDP i. e. purified fragments X, Y, or FbgDP generated in vitro in plasma by streptokinase treatment. This indicates that the assay is specific for fibrin degradation products.We have successfully applied this assay to the plasma of patients with a variety of diseased states. In combination with the assay previously developed by us for FbgDP and for the total amount of FbgDP + FbDP (TDP) in plasma, we are now able to study the composition of TDP in patients plasma in terms of FbgDP and FbDP.


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