A fully automated solid-phase radioimmunoassay for triiodothyronine.

1977 ◽  
Vol 23 (5) ◽  
pp. 851-854 ◽  
Author(s):  
N E Rugg ◽  
M J Hasler ◽  
R E Bjornsen ◽  
K Painter
Keyword(s):  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
Solid Phase ◽  
Correlation Coefficients ◽  
Ammonium Sulfate Precipitation ◽  
Men And Women ◽  
Coefficients Of Variation ◽  
Analytical Recovery ◽  
Solid Phase Radioimmunoassay ◽  
Normal Men

Abstract We report the first solid-phase radioimmunoassay for triiodothyronine with use of antibody-coated tubes, which is designed specifically for the fully automated radioimmunoassay instrument, Micromedic Systems' "Concept 4 Automatic Radioassay." Antisera to triiodothyropropionic acid/bovine serum albumin were raised in rabbits, purified by ammonium sulfate precipitation, and coated onto polypropylene tubes. Analytical recovery of exogenous triiodothyronine added to sera from normal men and women and pregnant women was quantitative. Intra-assay and inter-assay coefficients of variation are 4-5 and 6-9%, respectively. Correlation coefficients (r) for comparison of sample values with those obtained by three commercial laboratories were 0.95, 0.84, and 0.91. The sensitivity of the assay is 0.5 microng/liter. The assay can be performed either manually or be fully automated on the "Concept 4."

A solid-phase enzymeimmunoassay for the determination of progesterone in bovine, porcine and ovine plasma

1995 ◽  
Vol 75 (4) ◽  
pp. 525-529 ◽  
Author(s):  
R. P. Del Vecchio ◽  
W. D. Sutherland ◽  
M. L. Connor
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Petroleum Ether ◽  
Bovine Serum ◽  
Solid Phase ◽  
Plasma Samples ◽  
Assay Sensitivity ◽  
Coefficients Of Variation ◽  
Rabbit Igg

The purpose of this project was to develop a valid quantitative enzymeimmunoassay (EIA) for progesterone in blood plasma of cattle, pigs and sheep. Rabbit anti-progesterone, mouse monoclonal anti-rabbit IgG, authentic progesterone, and acetylcholine esterase bound covalently to progesterone were the principal reagents used to develop the EIA. Ninety-six well microliter plates were coated with mouse monoclonal anti-rabbit IgG and saturated with bovine serum albumin before use. Rabbit anti-progesterone was diluted to a working dilution of 1:2.0 × 106. Standard curves were linear and ranged from 1.56 to 400 pg of progesterone per well which allowed for the measurement of 0.03125 to 8.0 ng mL−1. Assay sensitivity averaged 1.56 pg well−1. Progesterone was extracted from plasma samples with petroleum ether. Plasma samples (n = 3 or 4 from each species) with unknown amounts of progesterone that were extracted and serially diluted with EIA buffer did not deviate from parallelism with progesterone standard curves in buffer. The correlation between EIA and radioimmunoassay (RIA) measurements of progesterone in the same plasma samples was high (P < 0.0001) for all three species (r = 0.96 for bovine; r = 0.96 for porcine; r = 0.94 for ovine). The regression of EIA data on RIA data produced the following equations:[Formula: see text]The intra- and inter-assay coefficients of variation were 5.4 and 10.6% for bovine, 5.8 and 11.0% for porcine and, 6.1 and 12.3% for ovine, respectively. These data show that this EIA is a valid and reliable memod for quantitating progesterone in extracts of bovine, porcine and ovine plasma. Key words: Enzymeimmunoassay, progesterone, plasma, bovine, porcine, ovine

A more specific, liquid-chromatographic method for free cortisol in urine.

1982 ◽  
Vol 28 (12) ◽  
pp. 2418-2420 ◽  
Author(s):  
E Canalis ◽  
G E Reardon ◽  
A M Caldarella
Keyword(s):  
Present Method ◽  
Chromatographic Method ◽  
Urinary Cortisol ◽  
Men And Women ◽  
Coefficients Of Variation ◽  
Liquid Chromatographic Method ◽  
Analytical Recovery ◽  
Free Cortisol ◽  
Normal Men ◽  
Liquid Chromatographic

Abstract Currently used assays for urinary cortisol reportedly overestimate it, owing to cross-reacting substances. We describe here a method for separating and measuring by liquid chromatography cortisol extracted from urine. The method is specific for cortisol and as little as 5 ng per sample can be measured. Mean analytical recovery of added cortisol was 98.8% (SD 6.1%) and the coefficients of variation ranged from 3.1 to 4.7% (within-day) and from 7.1 to 14% (between-day). Mean (and SD) urinary excretion of cortisol for 45 normal men and women was 20.1 (SD 7.6) micrograms/24 h; for 29 children it was 14.1 (SD 6.0) micrograms/24 h. Results by radioimmunoassay were 1.4- to 4.3-fold greater than by this method, and results of the two assays did not correlate well (r = 0.59, p less than 0.01). We consider the present method to be a practical and specific assay for three cortisol in urine.

A solid-phase radioimmunoassay for vitamin B12 in serum, with use of radioiodinated tyrosine methyl ester of vitamin B12.

1978 ◽  
Vol 24 (3) ◽  
pp. 460-465 ◽  
Author(s):  
D B Endres ◽  
K Painter ◽  
G D Niswender
Keyword(s):  
Methyl Ester ◽  
Bovine Serum Albumin ◽  
Vitamin B12 ◽  
Bovine Serum ◽  
Solid Phase ◽  
Radioactive Tracer ◽  
Factors Affecting ◽  
Denatured Protein ◽  
Solid Phase Radioimmunoassay ◽  
First Time

Abstract Although radioassays for vitamin B12 with use of any of several binding proteins have been available for many years, a radioimmunoassay for B12 has not been reported. We describe here such a radioimmunoassay, incorporating, for the first time, a radioiodinated tyrosine methyl ester of B12 as the radioactive tracer. Polypropylene tubes are coated with antiserum raised in a rabbit against B12/bovine serum albumin to simplify the separation of bound and free radioactivity. Factors affecting the preparation of coated tubes are described. The assay is accurate, sensitive, precise, and specific for vitamin B12. Accuracy of the assay is unaffected by the presence of denatured protein. The advantages of this radioimmunoassay over conventional radioassays are discussed.

Solid-phase radioimmunoassay of human complement fragment C5a.

1988 ◽  
Vol 34 (6) ◽  
pp. 1018-1021 ◽  
Author(s):  
S Chang ◽  
A Leo-Mensah ◽  
J Campbell ◽  
M Stastny ◽  
R A Patrick
Keyword(s):  
Incubation Time ◽  
Solid Phase ◽  
Biological Fluids ◽  
Reference Method ◽  
Human Complement ◽  
Analytical Recovery ◽  
Solid Phase Radioimmunoassay

Abstract In this competitive RIA for determining concentrations of human C5a in biological fluids and in buffers, labeled C5a and sample are allowed to compete for binding to a limited amount of goat antibody to human C5a in solution. Free and bound tracer are then separated by a second antibody (rabbit anti-goat IgG) immobilized on paramagnetic particles. Total incubation time for this assay is 70 min. Sensitivity, precision, and analytical recovery of this assay compare well with those of a reference method.

scholarly journals THE NATURE OF ANTIGEN-ANTIBODY COMPLEXES FORMED IN RABBITS DURING AN IMMUNE RESPONSE TO BOVINE SERUM ALBUMIN

1958 ◽  
Vol 107 (5) ◽  
pp. 653-663 ◽  
Author(s):  
William O. Weigle
Keyword(s):  
Immune Response ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Ammonium Sulfate ◽  
Bovine Serum ◽  
Ammonium Sulfate Precipitation ◽  
Antibody Synthesis ◽  
Circulating Antigen ◽  
Antigen Antibody

The immune elimination of soluble BSA, following an intravenous injection, is accompanied by the appearance of circulating antigen-antibody complexes. The pattern of the appearance of circulating antigen-antibody complexes and the immune elimination of antigen probably depends on the amount of antigen injected, the rate of antibody synthesis, and perhaps, the quality of antibody produced. There is no relationship between the I* antigen-antibody complexes detected during the immune response in rabbits by ammonium sulfate precipitation and the material precipitated from immune sera as a result of treatment with alkali. Alkali-precipitable material present in the serum of rabbits at a time when I* antigen is also present contain at most only traces of the antigen.

Rapid radioimmunoassay of total urinary estriol.

1976 ◽  
Vol 22 (5) ◽  
pp. 611-615 ◽  
Author(s):  
D W Anderson ◽  
U Goebelsmann
Keyword(s):  
Urinary Tract ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Ammonium Sulfate ◽  
Bovine Serum ◽  
Room Temperature ◽  
Urine Specimen ◽  
Ammonium Sulfate Precipitation ◽  
In Pregnancy

Abstract We describe a radioimmunoassay for total urinary estriol in pregnancy. A 25-mul aliquot of the urine specimen is acid hydrolyzed, neutralized, and diluted before assay. We use rabbit antisera against estriol-6-(O-carboxymethyl)oxime/bovine serum albumin and ammonium sulfate precipitation at room temperature. Results are unaffected by glucose or methenamine mandelate, a urinary tract antiseptic. Using semi-automatic pipetting equipment, one laboratory technologist can complete 50 assays within 8 h. This technique is both reliable and convenient and should decrease the expense of routine estriol assays.

Antibodies to Testosterone-3-Bovine Serum Albumin, Applied to Assay of Serum 17β-ol Androgens

1971 ◽  
Vol 17 (7) ◽  
pp. 581-584 ◽  
Author(s):  
Jeffrey C Chen ◽  
Elinor M Zorn ◽  
Marvin C Hallberg ◽  
Ralph G Wieland
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Binding Assay ◽  
Virgin Female ◽  
Competitive Binding ◽  
Competitive Binding Assay ◽  
Menstruating Women ◽  
Normal Men ◽  
Mean Concentration

Abstract Testosterone was conjugated to bovine serum albumin at carbon 3, and antibodies to this conjugate were developed in virgin female rabbits. This antiserum, diluted 1:3200, was used to measure testosterone. With tritiated testosterone and anti-rabbit γ-globulin as the second antibody, a doseresponse curve was obtained for doses in the range 0.1-10 ng. The antibody cross-reacts extensively with dihydrotestosterone, so that dihydrotestosterone is measured with testosterone in extracted but unchromatographed serum. Results for pooled serum from men, as measured repeatedly by radioimmunoassay and competitive binding assay for testosterone, agreed well. Seventeen normal men had serum 17β-ol androgen concentrations of 667 ng/100 ml (SD, ±248); seventeen normal, menstruating women had a mean concentration of 69 ng/100 ml (SD, ±18). Values for 17β-ol androgens in adolescent males agreed reasonably with testosterone values reported by other investigators. Values are presented for patients with various androgenic disorders. This procedure is easy; many samples can be processed at one time.

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