Determination of picomolar equilibrium dissociation constants in solution by enzyme-linked immunosorbent assay with fluorescence detection

2005 ◽  
Vol 347 (1) ◽  
pp. 159-161 ◽  
Author(s):  
Katrina High ◽  
Yuan Meng ◽  
Michael W. Washabaugh ◽  
Qinjian Zhao
Keyword(s):  
Immunosorbent Assay ◽  
Fluorescence Detection ◽  
Dissociation Constants ◽  
Enzyme Linked Immunosorbent Assay ◽  
Equilibrium Dissociation Constants ◽  
Equilibrium Dissociation

scholarly journals Circular dichroism determination of class I MHC-peptide equilibrium dissociation constants

1997 ◽  
Vol 6 (8) ◽  
pp. 1771-1773 ◽  
Author(s):  
Chantal S. Morgan ◽  
James M. Holton ◽  
Barry D. Olafson ◽  
Pamela J. Bjorkman ◽  
Stephen L. Mayo
Keyword(s):  
Circular Dichroism ◽  
Dissociation Constants ◽  
Class I ◽  
Class I Mhc ◽  
Equilibrium Dissociation Constants ◽  
Equilibrium Dissociation ◽  
Circular Dichroïsm

Determination of Equilibrium Dissociation Constants

2009 ◽  
pp. 503-524 ◽  
Author(s):  
Robin E. Ernst ◽  
Katrina N. High ◽  
Tom R. Glass ◽  
Qinjian Zhao
Keyword(s):  
Dissociation Constants ◽  
Equilibrium Dissociation Constants ◽  
Equilibrium Dissociation

SENSITIVE AND SPECIFIC ENZYME-LINKED IMMUNOSORBENT ASSAY FOR UROKINASE-TYPE PLASMINOGEN ACTIVATOR IN HUMAN PLASMA

1987 ◽  
Author(s):  
J Grøndahl-HANSEN ◽  
N Agerlin ◽  
L S Nielsen ◽  
K Danø
Keyword(s):  
Plasminogen Activator ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mean Values ◽  
Amino Terminal ◽  
Type Plasminogen Activator ◽  
Healthy Donors ◽  
Urokinase Type Plasminogen Activator ◽  
The Mean

An enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of human urokinase-type plasminogen activator (u-PA) in plasma and serum. Microtiter plates were coated with a monoclonal antibody and incubated with standard or sample. Bound u-PA was quantitated with polyclonal antibodies conjugated with biotin, followed by avidin-peroxidase. The assay was 10-fold as sensitive as other previously reported ELISAs, the detection limit being approximately 1 pg of u-PA in a volume of 100 μl with a linear dose-response up to 15 pg of u-PA. The assay detected active u-PA and its inactive proenzyme form equally well and the recovery of both forms was higher than 90% in plasma. A variety of structurally related proteins, including t-PA, were tested, but no reaction with proteins other than u-PA and its amino-terminal degradation product were observed. The intra-assay and inter-assay coefficients of variation for determination of u-PA in plasma were 7.6% and 8.4%, respectively. The assay was equally applicable to serum. The values obtained with plasma and serum were similar, and the results were not affected by small variations in the preparation of the samples. The ELISA was used to measure the concentration of u-PA in plasma from 34 healthy donors. The mean values for u-PA in plasma from healthy donors was 1.1 ng/ml ± 0.3 ng/ml (SD) (range 0.6 - 1.5 ng/ml). No significant differences were found between men and women and no correlation between u-PA concentration and age could be demonstrated.The mean u-PA concentration in plasma from healthy donors obtained in this study is substantially lower than that reported by others. This might be due to different methods of determination of the protein content of the standard preparations or to differences in the specificity of the assays.

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