Direct 125I-radioligand assays for serum progesterone compared with assays involving extraction of serum.

1982 ◽  
Vol 28 (6) ◽  
pp. 1314-1318 ◽  
Author(s):  
W A Ratcliffe ◽  
J E Corrie ◽  
A H Dalziel ◽  
J S Macpherson
Keyword(s):  
Human Serum ◽  
Binding Proteins ◽  
Serum Progesterone ◽  
Double Antibody ◽  
Analytical Recovery ◽  
Conventional Methods

Abstract We compared two direct radioimmunoassays for progesterone in 50 microL of unextracted serum or plasma with assays involving extraction of serum. The direct assays include the use of either danazol at pH 7.4 or 8-anilino-1-naphthalenesulfonic acid at pH 4.0 to displace progesterone from serum binding-proteins. Progesterone is then assayed by using an antiserum to a progesterone 11 alpha hemisuccinyl conjugate and the radioligand 125I-labeled progesterone 11 alpha-glucuronyl tyramine, with separation by double-antibody techniques. Direct assays with either displacing agent gave good analytical recovery of progesterone added to human serum, and progesterone values for patients' specimens correlated well (r greater than 0.96) with results of assays involving extraction of serum. Precision was similar with each displacing agent over the working range 2.5-100 nmol/L and superior to that of extraction assays. We conclude that these direct assays of progesterone are analytically valid and more robust, precise, and technically convenient than many conventional methods involving extraction of serum.

Simultaneous radioimmunoassay of thyrotropin and thyroxine in human serum.

1977 ◽  
Vol 23 (9) ◽  
pp. 1644-1647 ◽  
Author(s):  
M K Bluett ◽  
E O Reiter ◽  
G E Duckett ◽  
A W Root
Keyword(s):  
Human Serum ◽  
Congenital Hypothyroidism ◽  
Diagnosis And Treatment ◽  
Rapid Evaluation ◽  
Dual Isotope ◽  
Screening Programs ◽  
Double Antibody ◽  
Specimen Collection ◽  
Analytical Recovery

Abstract The importance of early diagnois and treatment of congenital hypothyroidism has been well established, and several screening programs have been undertaken to detect neonates with this disorder by measurement of concentrations of thyrotropin or thyroxine in the serum. However, measurement of either hormone alone may fail to identify all affected patients. Accordingly, we have established a simultaneous double-antibody, dual-isotope radioimmunoassay for both. Sensitivity, slope, analytical recovery, and precision characteristics of the simultaneous assay do not differ from those of each assay performed separately. Values for the two analyses in the single and simultaneous assays correlate well (r = 0.951 for thyroxine, 0.983 for thyrotropin). This assay system permits determination of both hormones within 72 h after specimen collection and thus should allow more rapid evaluation, diagnosis, and treatment of infants with congenital hypothyroidism.

Some Critical Factors in Double Antibody Radioimmunoassay Systems Utilizing Sheep Anti-rabbit Precipitating Sera for Measurement of Human Serum LH, FSH and HGH

1969 ◽  
Vol 29 (7) ◽  
pp. 948-956 ◽  
Author(s):  
I. M. BURR ◽  
D. B. GRANT ◽  
P. C. SIZONENKO ◽  
S. L. KAPLAN ◽  
M. M. GRUMBACH
Keyword(s):  
Human Serum ◽  
Critical Factors ◽  
Double Antibody ◽  
Serum Lh

Activin-binding proteins in human serum and follicular fluid.

1992 ◽  
Vol 74 (6) ◽  
pp. 1320-1324
Author(s):  
A L Schneyer ◽  
D A O'Neil ◽  
W F Crowley
Keyword(s):  
Human Serum ◽  
Follicular Fluid ◽  
Binding Proteins

scholarly journals PROPERTIES OF VITAMIN B12-BINDING PROTEINS IN HUMAN SERUM

1959 ◽  
Vol 5 (1) ◽  
pp. 35-43 ◽  
Author(s):  
NOBUO KATO
Keyword(s):  
Vitamin B12 ◽  
Human Serum ◽  
Binding Proteins

Solid-phase enzymoimmunoassay for osteocalcin in human serum or plasma, with use of a monoclonal antibody.

1989 ◽  
Vol 35 (10) ◽  
pp. 2087-2092 ◽  
Author(s):  
M J Power ◽  
P F Fottrell
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Horseradish Peroxidase ◽  
Human Serum ◽  
Solid Phase ◽  
Sample Volume ◽  
Microtiter Plates ◽  
Analytical Recovery ◽  
Peroxidase Conjugate

Abstract In this solid-phase enzymoimmunoassay on microtiter plates for osteocalcin in serum or plasma, we use an osteocalcin-horseradish-peroxidase conjugate and a monoclonal antibody raised against bovine osteocalcin. We thoroughly standardized the assay for measurement of osteocalcin in both serum and plasma, demonstrating independence of sample volume, and determining the analytical recovery and within-and between-assay CVs. The detection limit was between 0.6 and 1.1 micrograms/L and the ED50 was 16 micrograms/L for a 5-microL sample volume. The intra-assay CV over the range 3 to 74 micrograms/L was less than or equal to 15%. The interassay CV over the range 3.6 to 46 micrograms/L was less than or equal to 16%. Results by this assay and by an in-house radioimmunoassay in which the same monoclonal antibody was used correlated well (r2 = 0.948). Osteocalcin concentrations in serum and plasma as measured with the present assay agreed well with published values.

Improved radioimmunoassay of urinary estriol.

1979 ◽  
Vol 25 (1) ◽  
pp. 99-102 ◽  
Author(s):  
M J Jawad ◽  
E A Wilson ◽  
H L Kincaid
Keyword(s):  
Incubation Time ◽  
Colorimetric Method ◽  
Detection Range ◽  
Standard Curve ◽  
Double Antibody ◽  
Analytical Recovery ◽  
Urinary Glucose

Abstract We report a rapid double-antibody radioimmunoassay for urinary estriol. Advantages over other current methods include: (a) 30-min hydrolysis; (b) total incubation time, 55 min; (c) assay unaffected by urinary glucose; (d) no degradation of estriol evident during hydrolysis; (e) superior (85%) analytical recovery of estriol conjugates; (f) linear standard curve by logit-log extrapolation; (g) good correlation (r = 0.83) with total estrogen determination by a generally accepted colorimetric method; (h) only 20 muL of urine required; and (i) the detection range is 1.9 to 100.5 mg/24-h urine.

A coupled-enzyme equilibrium method for measuring urea in serum: optimization and evaluation of the AACC study group on urea candidate reference method.

1980 ◽  
Vol 26 (7) ◽  
pp. 816-826 ◽  
Author(s):  
E J Sampson ◽  
M A Baird ◽  
C A Burtis ◽  
E M Smith ◽  
D L Witte ◽  
...  
Keyword(s):  
Human Serum ◽  
Glutamate Dehydrogenase ◽  
Reference Method ◽  
Linear Range ◽  
Study Group ◽  
Reaction Conditions ◽  
Equilibrium Method ◽  
Analytical Recovery

Abstract We describe a coupled-enzyme equilibrium method for measuring urea in serum, which is performed on supernates prepared by treating each specimen with Ba(OH)2 and ZnSO4 (Somogyi reagent). Analytical recovery of [14C]urea added to a variety of matrices was essentially complete (mean, 100.6%) for the supernates after precipitation. Nine variables were univariately examined in arriving at the reaction conditions for the method: glutamate dehydrogenase, urease, 2-oxoglutarate, ADP, Tris . HCI, NADH, EDTA, pH, and temperature. The reagent is stable for at least 48 days at--20 degrees C and for 23 days at 4 degrees C. Mean analytical recovery of urea (14 mmol/L) added to seven different specimens (three different matrices) was 100.8%. The analytical linear range of the method extends to 30 mmol of urea per liter. Of 22 potential interferents, only bilirubin at 1 mmol/L (580 mg/L), hemoglobin at 10 g/L, and hydroxyurea at 6 mmol/L showed more than 2% interference. We discuss precision and effects of specimen dilution, and compare results for 100 human serum specimens with those measured for the same specimens with four other urea methods. We examined the effects of measuring a blank, consisting of sample and reagent without urease, with each specimen.

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