Competitive protein-binding radioassay of 24,25-dihydroxyvitamin D in sera from normal and anephric subjects

1977 ◽  
Vol 182 (2) ◽  
pp. 390-395 ◽  
Author(s):  
John G. Haddad ◽  
Chong Min ◽  
Mike Mendelsohn ◽  
Eduardo Slatopolsky ◽  
Theodore J. Hahn
1980 ◽  
Vol 201 (1) ◽  
pp. 277-285 ◽  
Author(s):  
John P. Mallon ◽  
James G. Hamilton ◽  
Cheryl Nauss-Karol ◽  
Robert J. Karol ◽  
Constance J. Ashley ◽  
...  

1981 ◽  
Vol 27 (3) ◽  
pp. 444-450 ◽  
Author(s):  
M J Jongen ◽  
W J van der Vijgh ◽  
H J Willems ◽  
J C Netelenbos

Abstract 1,25-Dihydroxyvitamin D in plasma is measured by competitive protein-binding assay after isolation from plasma. The present method is improved, as compared with those hitherto described, with regard to receptor preparation, isolation of vitamin D metabolites from plasma, and procedure for the competitive protein binding. Receptor preparation from healthy rather than rachitic chicks, together with a fast isolation procedure, results in a high yield of active receptor protein: 12 animals provide receptor for about 4000 incubations. No loss of binding activity was observed during one year. 1,25-Dihydroxyvitamin D is isolated from plasma by a single extraction and a one-step chromatographic purification. Analytical recovery for the entire procedure averaged 78.1% (SD 4.7%). Other vitamin D metabolites (25-hydroxyvitamin D and 24,25-dihydroxyvitamin D) can also be separated with this procedure. The main features of the modified binding assay are the use of a stabilized cytosol receptor and dextran-coated charcoal instead of polyethylene glycol. The smallest detectable amount in the binding assay is 1-2 pg (2.4-4.8 fmol). Intra- and interassay CVs are 7.0% and 4.8%, respectively. 1,25-Dihydroxyvitamin D concentrations in plasma of 20 healthy subjects averaged 51.7 (SD 10.8) ng/L [124 (SD 26) pmol/L]. Three anephric patients showed values of 3, 6, and 7 ng/L (7, 14, and 17 pmol/L).


1978 ◽  
Vol 25 (5) ◽  
pp. 431-436 ◽  
Author(s):  
SHIGEHARU DOKOH ◽  
RIKUSHI MORITA ◽  
MASAO FUKUNAGA ◽  
ITSUO YAMAMOTO ◽  
KANJI TORIZUKA

1970 ◽  
Vol 64 (4) ◽  
pp. 630-636 ◽  
Author(s):  
Stephen C. Thorson ◽  
Ronald Tsujikawa ◽  
James L. Brown ◽  
Robert T. Morrison ◽  
Hamish W. McIntosh

ABSTRACT Serum thyroxine concentrations were determined in 66 euthyroid, 30 hyperthyroid and 13 hypothyroid patients using both the established Murphy method and a simplified method of competitive protein binding analysis. A diagnosis compatibility of 96% was found with both methods indicating that the simplified method has comparable clinical application as an initial screen of thyroid status.


1970 ◽  
Vol 63 (2) ◽  
pp. 225-241 ◽  
Author(s):  
B. D. Reeves ◽  
M. L. A. de Souza ◽  
I. E. Thompson ◽  
E. Diczfalusy

ABSTRACT An improved method for the assay of plasma progesterone by competitive protein binding is described. The improvement is based upon rigorous control of the variables, the compensation for and standardisation of interfering factors inherent in the method and the use of a human corticosteroid binding globulin, that meets the requirements for sensitivity at levels of 1.0 ng of progesterone and below. The assessment of the reliability of the individual steps in the method as well as that of the complete method is presented. The sensitivity of the method is around 0.2 ng progesterone per ml plasma. Accuracy was measured by adding progesterone in amounts ranging from 0.0 to 1.0 ng to 1.0 ml plasma. There was a linear relationship between the progesterone added and recovered throughout the entire range of values, with a coefficient of correlation (r) of 0.94. Of 52 related steroids tested, none was found which would remain associated with progesterone following extraction and purification and which would also compete with progesterone for binding sites.


1970 ◽  
Vol 65 (1_Suppl) ◽  
pp. S61-S78 ◽  
Author(s):  
Billy D. Reeves ◽  
David W. Calhoun

ABSTRACT This communication is an attempt to delineate and define reliability criteria for saturation analysis of steroids by competitive protein binding assay. The discussion of these criteria evolved from three major considerations of assay method that help to place the ultimate criterion of accuracy in proper perspective. These major considerations are: 1) the measurement system, 2) the assay design and 3) the calculations and statistical control. Such an approach permits an evaluation, both relative and absolute, for a single method or for multiple methods.


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