DEVELOPMENT OF A RADIOIMMUNOASSAY FOR 5α-ANDROST-16-EN-3-ONE IN PIG PERIPHERAL PLASMA

1974 ◽  
Vol 76 (2) ◽  
pp. 377-387 ◽  
Author(s):  
Ø. Andresen
Keyword(s):  
Regression Analysis ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Cross Reaction ◽  
Plasma Samples ◽  
Standard Curve ◽  
Peripheral Plasma ◽  
Accuracy Study ◽  
The Mean

ABSTRACT A radioimmunoassay (RIA) for 5α-androst-16-en-3-one (5α-androstenone) in peripheral plasma from pigs has been developed. Antibodies against 5α-androstenone conjugated through C-3 to bovine serum albumin (BSA) were produced in rabbits. The antiplasma used in this study show a cross-reaction of 100% with 4,16-androstadien-3-one (androstadienone), 5.3 % with 5α-androst-16-en-3β-ol (an-β), 3.3 % with 5α-androst-16-en-3α-ol (an-α), 2.6 % with 4-androstene-3,17-dione (androstenedione) and 1.4% with 5,16-androstadien-3β-ol (andien-β). The standard curve plotted as log picogram (pg) unlabelled 5α-androstenone against counts per minute (cpm) bound radioactive steroid was almost linear from 50 to 800 pg 5α-androstenone. Regression analysis of the data from the accuracy study gave the curve y = 0.99 x+ 203. The precision and sensitivity of the method were satisfactory. In plasma samples from sows and castrated male pigs the mean 5α-androstenone found was 1.1 ng/ml. In plasma samples from boars from 1.2 to 54.1 ng/ml was found.

RADIOIMMUNOASSAY OF OESTRONE AND OESTRADIOL IN THE PERIPHERAL PLASMA OF THE DOMESTIC FOWL IN VARIOUS PHYSIOLOGICAL STATES AND OF HYPOPHYSECTOMIZED AND OVARIECTOMIZED FOWLS

1974 ◽  
Vol 75 (1) ◽  
pp. 133-140 ◽  
Author(s):  
B. E. Senior
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Diethyl Ether ◽  
Bovine Serum ◽  
Domestic Fowl ◽  
Laying Hens ◽  
Peripheral Plasma ◽  
The Mean ◽  
Precision And Accuracy ◽  
Chicken Ovary

ABSTRACT A radioimmunoassay was developed to measure the levels of oestrone and oestradiol in 0.5–1.0 ml of domestic fowl peripheral plasma. The oestrogens were extracted with diethyl ether, chromatographed on columns of Sephadex LH-20 and assayed with an antiserum prepared against oestradiol-17β-succinyl-bovine serum albumin using a 17 h incubation at 4°C. The specificity, sensitivity, precision and accuracy of the assays were satisfactory. Oestrogen concentrations were determined in the plasma of birds in various reproductive states. In laying hens the ranges of oestrone and oestradiol were 12–190 pg/ml and 29–327 pg/ml respectively. Levels in immature birds, in adult cockerels and in an ovariectomized hen were barely detectable. The mean concentrations of oestrone and oestradiol in the plasma of four non-laying hens (55 pg/ml and 72 pg/ml respectively) and one partially ovariectomized hen (71 pg/ml and 134 pg/ml respectively) were well within the range for laying hens. It is evident that the large, yolk-filled follicles are not the only source of oestrogens in the chicken ovary.

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

Screening radioimmunoassay for aldosterone in preheated plasma without extraction and chromatography.

1980 ◽  
Vol 26 (1) ◽  
pp. 41-45
Author(s):  
T M Connolly ◽  
L Tibor ◽  
K H Gless ◽  
P Vecsei
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Binding Proteins ◽  
Equilibrium Dialysis ◽  
Screening Method ◽  
Plasma Aldosterone ◽  
Plasma Samples ◽  
Bovine Serum Albumin Conjugate ◽  
Native Plasma

Abstract We directly estimated plasma aldosterone radioimmunologically with use of an antiserum raised against an aldosterone-3-oxime/bovine serum albumin conjugate, the estimation being on samples with and without heating (60 degrees C), and diluted and undiluted. Values so obtained were compared with those by radioimmunoassay after extraction and chromatography. The correlation--even negative values were obtained--was poorest when the steroid was directly estimated in nonheated, undiluted plasma. Correlations were best (r = 0.918) for preheated and diluted native plasma, and the interassay CV was 9.8% (n = 57). However, there were some extraordinarily high values. After equilibrium dialysis of native and preheated (60 degrees C) plasma (15 plasma samples), the percentages of apparent free aldosterone and cortisol increased from 51.4 +/- 2.6% (SEM) to 64.3 +/- 1.6% and from 11.5 +/- 2.2% to 61.1 +/- 1%, respectively. We conclude that aldosterone-binding proteins play a role in direct radioimmunoassays of aldosterone in plasma, but by heating (with or without diluting) the plasma, direct assay can be used as a simple, fast, and inexpensive screening method.

scholarly journals Radioimmunoassay of colchicine

1975 ◽  
Vol 151 (2) ◽  
pp. 413-415 ◽  
Author(s):  
C Boudene ◽  
F Duprey ◽  
C Bohuon
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Biological Fluids ◽  
Cross Reaction ◽  
Colchicine Derivatives

Antibodies to cholchicine were raised in rabbits after injection of N-desacetylthiocolchicine conjugated to bovine serum albumin. Antisera diluted 1:100 bound 50% of 1.6ng of [3H]colchicine. Cross-reaction of the antisera with colchicine derivatives and other drugs was measured and a radioimmunoassay of colchicine in biological fluids was proposed.

Oxygen concentration and protein source affect the development of preimplantation goat embryos in vitro

1991 ◽  
Vol 3 (5) ◽  
pp. 601 ◽  
Author(s):  
PA Batt ◽  
DK Gardner ◽  
AW Cameron
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Oxygen Concentration ◽  
Bovine Serum ◽  
Calf Serum ◽  
Cell Number ◽  
The Mean ◽  
Number Of Cells

The effect of oxygen concentration and the source of protein in culture medium on the development of 2- to 4-cell goat embryos in vitro was investigated. Embryos were collected from superovulated Angora-Cashmere-cross goats 48 h after ovulation and cultured for 6 days in synthetic oviduct fluid (SOF) medium under one of two oxygen concentrations (20% or 7%) and in the presence of one of five protein sources; Miles bovine serum albumin (Miles BSA), Commonwealth Serum Laboratory bovine serum albumin (CSL BSA), goat serum (GS), fetal calf serum (FCS) and human serum (HS). In the presence of 20% oxygen the percentage of embryos reaching the expanded and/or hatched blastocyst stage in SOF medium containing Miles BSA was 29%, with a mean cell number per embryo of 28.1 +/- 6.0 (+/- s.e.m.). Use of an oxygen concentration of 7% significantly increased the percentage of embryos reaching this stage (80%, P less than 0.01) and the mean number of cells per embryo (65.3 +/- 8.2, P less than 0.01). The mean number of cells of the early-cleavage-stage embryos was significantly lower when the medium contained CSL BSA, GS or FCS (42.7 +/- 5.6, 29.0 +/- 6.1 and 21.3 +/- 3.2, respectively) than with Miles BSA (92.8 +/- 6.4) or HS (104.8 +/- 17.2) (P less than 0.01). Under 7% oxygen and with Miles BSA or HS, embryos were morphologically comparable to those developed in vivo, but the mean cell numbers in vitro were only approximately half those obtained in vivo.

Successful Reversal of Diabetes by Single Donor Isologous Islet Transplantation in a Mouse Model

1997 ◽  
Vol 6 (4) ◽  
pp. 429-430 ◽  
Author(s):  
C. K. Leow ◽  
D. W. R. Gray ◽  
P. J. Morris
Keyword(s):  
Bovine Serum Albumin ◽  
Mouse Model ◽  
Serum Albumin ◽  
Islet Transplantation ◽  
Bovine Serum ◽  
High Yield ◽  
Renal Capsule ◽  
Single Donor ◽  
Mouse Islets ◽  
The Mean

A method for isolating mouse islets which consistently gives a high yield with good purity is described. Using a bovine serum albumin gradient, the mean yield of islets per pancreas is 425 (SEM ± 15) with a consistent purity of over 90%. Single donor to single recipient of islets transplanted under the renal capsule restores normoglycemia in the diabetic recipients within 2 to 5 days of transplantation.

Microencapsulated antibodies in radioimmunoassay. III. Determination of cortisol.

1980 ◽  
Vol 26 (5) ◽  
pp. 633-634 ◽  
Author(s):  
R W Bordens ◽  
E P Halpern
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Plasma Cortisol ◽  
Total Plasma ◽  
Competitive Reaction ◽  
Large Molecules ◽  
Standard Curve ◽  
Analytical Recovery

Abstract We describe a method for directly measuring total plasma cortisol by use of a microencapsulated antibody. The antibody microcapsules were incubated with plasma and 125I-labeled cortisol in a competitive reaction at 37 degrees C for 15 min, then separated by centrifugation; the radioactivity of the pellet was counted for 1 min. Analytical recovery of cortisol from three plasma pools was 97.6% (SD 8.5%) and was unaffected by triglyceridemia, hemolysis, or icterus. The standard curve was linear to 500 microgram/L and the sensitivity was 10 microgram/l. Recovery of cortisol added as the hemisuccinate was 102% (SD 10.6%), whereas that of the conjugate cortisol hemisuccinate/bovine serum albumin was 4.3% (SD 3.5%). This confirmed that the microcapsule excludes large molecules. The within-day CV for two control pools was 9.8 and 12.8%, the day-to-day variation 13.4 and 13.8%.

scholarly journals PERSISTENCE OF HAPTEN-ANTIBODY COMPLEXES IN THE CIRCULATION OF IMMUNIZED ANIMALS AFTER A SINGLE INTRAVENOUS INJECTION OF HAPTEN

1974 ◽  
Vol 139 (2) ◽  
pp. 278-294 ◽  
Author(s):  
Donald H. Schmidt ◽  
Bette M. Kaufman ◽  
Vincent P. Butler
Keyword(s):  
Molecular Weight ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Half Life ◽  
Bovine Serum ◽  
Single Injection ◽  
Low Molecular Weight ◽  
Serum Digoxin ◽  
Biological Half Life ◽  
The Mean

To study the fate of a low molecular weight antigen (hapten) in the circulation of animals whose sera contain antibodies specific for that low molecular weight antigen, a single injection of digoxin-3H (0.4 mg/kg) was administered intravenously to 18 rabbits. Thirteen animals (nine nonimmunized and four immunized with bovine serum albumin) served as control animals. In five rabbits which had been immunized with a digoxin-bovine serum albumin conjugate and whose sera contained digoxin-specific antibodies, the mean 12-h serum digoxin concentration was 8,300 ng/ml (control: 92 ng/ml) and the mean serum concentration 12 mo after the single injection of digoxin-3H was 85 ng/ml. In digoxin-immunized rabbits, less than 10% of the digoxin-3H was excreted in the first 10 days (control: 77% recovered in urine and feces) and the mean biological half-life of digoxin, as calculated from serum digoxin-3H disappearance curves, was 72 days (control: 3.4 days). In sera of digoxin-immunized rabbits, more than 90% of the circulating digoxin-3H was immunoglobulin bound, as determined by the double-antibody and dextran-coated charcoal methods. The serum disappearance rate of 125I-antidigoxin antibodies was similar in nonimmunized and in immunized animals and in the presence or absence of digoxin. It is concluded that the biological half-life of a hapten may be markedly prolonged when the hapten is bound to specific antibody. The persistence of antibody-hapten complexes in the circulation suggests that these complexes may not be deposited in tissues and raises the possibility that low molecular weight determinants may be capable of preventing or reversing the deposition of immune complexes, containing macromolecular antigens, in the tissues of experimental animals and man.

scholarly journals Production and properties of antisera to dexamethasone–protein conjugates

1973 ◽  
Vol 133 (2) ◽  
pp. 401-404 ◽  
Author(s):  
M. C. Dumasia ◽  
D. I. Chapman ◽  
M. S. Moss ◽  
C. O'Connor
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Cross Reaction ◽  
Protein Conjugates ◽  
The Cross

Antibodies to dexamethasone 21-hemisuccinate conjugated to bovine serum albumin were produced in rabbits. Antisera diluted 1:3000 bound 50% of 90pg of [1,2-3H]dexamethasone. The cross-reaction of the antisera with other synthetic and natural corticosteroids was measured.

Sign in / Sign up

Export Citation Format

Share Document