scholarly journals A radioimmunoassay for chicken avidin. Comparison with a [14C]biotin-binding method

1978 ◽  
Vol 175 (2) ◽  
pp. 685-690 ◽  
Author(s):  
M S Kulomaa ◽  
H A Elo ◽  
P J Tuohimaa
Keyword(s):  
Binding Sites ◽  
Solid Phase ◽  
Solid Matrix ◽  
Chicken Tissues ◽  
Standard Curve ◽  
Double Antibody ◽  
Liver And Kidney ◽  
Biotin Binding ◽  
Solid Phase Radioimmunoassay ◽  
Chloramine T

A double-antibody solid-phase radioimmunoassay for chicken avidin is reported. Avidin was labelled with 125I by the chloramine-T method. The bound and free avidin were separated with a second antibody bound to a solid matrix. In the logit-log scale the standard curve was linear from 1-2 to 100-200ng of avidin/ml. Cross-reaction of ovalbumin was less than 0.015%. Saturation of biotin-binding sites of avidin with an excess of biotin decreased radioimmunoassay values by about 15%. Recovery studies indicated that avidin can be assayed from all chicken tissues studied with radioimmunoassay, whereas the [14C]biotin/bentonite method gave poor recoveries for avidin in the liver and kidney. Radioimmunoassay and the [14C]biotin/bentonite method gave similar concentrations for oviduct avidin.

Double-antibody radioimmunoassay of thyroid microsomal antibody in serum.

1981 ◽  
Vol 27 (1) ◽  
pp. 39-42 ◽  
Author(s):  
V T Kung ◽  
P M Weber ◽  
L V dos Remedios
Keyword(s):  
Solid Phase ◽  
Sodium Deoxycholate ◽  
Published Data ◽  
Health Examination ◽  
Standard Curve ◽  
Double Antibody ◽  
Microsomal Antibody ◽  
Solid Phase Radioimmunoassay ◽  
Chloramine T ◽  
Antimicrosomal Antibody

Abstract We describe a double-antibody radioimmunoassay for antimicrosomal antibodies in serum. Antigen in centrifuged pellets of microsome is solubilized by treatment with sodium deoxycholate solution and radiolabeled (125I] by the Chloramine T method. A 10-microL aliquot of human serum or antimicrosomal antibody standard is incubated overnight at room temperature with 125I-labeled microsome. Bound radiolabeled microsome is separated by precipitation with goat antihuman IgG antiserum. The percentage of 125I-labeled microsome bound by each serum is calculated, and its concentration of antimicrosomal antibody interpolated from a standard curve. The prevalence of abnormal antimicrosomal antibody was 16% in a self-referred asymptomatic or not acutely ill group of patients having a periodic multiphasic health examination. Values for patients with certain thyroid diseases generally agreed with other published data. We conclude that our assay is sensitive, precise, and accurate, and is more efficient for routine measurement of antimicrosomal antibody than the currently used solid-phase radioimmunoassay.

scholarly journals A Rapid Solid-Phase Radioimmunoassay for Human Plasma Follicle-Stimulating Hormone

1980 ◽  
Vol 17 (1) ◽  
pp. 38-44
Author(s):  
AC Lovesey
Keyword(s):  
Human Plasma ◽  
High Precision ◽  
Reference Values ◽  
Plasma Levels ◽  
Solid Phase ◽  
Follicle Stimulating Hormone ◽  
Diagnosis And Management ◽  
Double Antibody ◽  
Solid Phase Radioimmunoassay ◽  
Stimulating Hormone

The measurement of plasma levels of human follicle-stimulating hormone (FSH) has proved to be of value for the study of the hypothalamic-hypophyseal-gonadal axis, greatly facilitating the diagnosis and management of problems relating to the menopause and infertility. In the present work a solid-phase radioimmunoassay for human FSH has been developed. This system is characterised by high precision, is economical, and is considerably faster to operate than conventional double antibody systems used in the hospital assay service. Reference values for plasma FSH in various endocrine states are recorded and discussed.

Basal levels of prolactin in goat blood measured throughout a 24-h period by a rapid double antibody–solid phase radioimmunoassay

1973 ◽  
Vol 40 (2) ◽  
pp. 235-245 ◽  
Author(s):  
I. C. Hart
Keyword(s):  
Solid Phase ◽  
Virgin Female ◽  
Double Antibody ◽  
Lactating Goats ◽  
Highly Sensitive ◽  
Solid Phase Radioimmunoassay ◽  
Circulating Levels

SummaryThe development of a rapid, highly sensitive double antibody–solid phase (DASP) radioimmunoassay for ovine/caprine prolactin is described. The assay was applied to the measurement of basal levels of prolactin in the blood of 4 anoestrous lactating female, 4 anoestrous virgin female and 4 castrated male goats. For several hours after milking there was a chronic gradually decreasing release of prolactin in the lactating goats, such that the circulating levels of prolactin were, for 14–16 h of the day, at a much higher level than those in the virgin female or castrated male goats. The basal levels of prolactin in the castrated male goats (87–200ng/ml) were somewhat higher than those in the virgin females (47–77ng/ml). The effects of feeding and stress on prolactin levels are discussed.

scholarly journals Assay of Australia Antigen Employing Double-Antibody and Solid-Phase Radioimmunoassay Techniques

Kanzo ◽  
1974 ◽  
Vol 15 (7) ◽  
pp. 427-432
Author(s):  
Shinichi KAKUMU ◽  
Shozo ITO ◽  
Sumihiko OKUYAMA ◽  
Seikichi INADA ◽  
Katsuhiko TAGO
Keyword(s):  
Solid Phase ◽  
Australia Antigen ◽  
Double Antibody ◽  
Solid Phase Radioimmunoassay

POTENCY ESTIMATES OF HUMAN GONADOTROPHINS BY A BIOASSAY AND THREE IMMUNOASSAY METHODS

1971 ◽  
Vol 67 (3) ◽  
pp. 417-433 ◽  
Author(s):  
C. Robyn ◽  
M. L'Hermite ◽  
P. Petrusz ◽  
E. Diczfalusy
Keyword(s):  
Biological Activity ◽  
Solid Phase ◽  
Haemagglutination Inhibition ◽  
Reproductive Organs ◽  
Male Rats ◽  
Equilibrium System ◽  
Antibody Technique ◽  
Double Antibody ◽  
Human Menopausal Gonadotrophin ◽  
Solid Phase Radioimmunoassay

ABSTRACT Human chorionic gonadotrophin (HCG), human menopausal gonadotrophin (HMG) and human hypophysial gonadotrophin (HHG) preparations were assayed by a bioassay and three immunoassay methods. The bioassay method was based on the increase in weight of the accessory reproductive organs of immature male rats. The immunoassays included haemagglutination inhibition, a radioimmunoassay using the double antibody technique and a solid phase radioimmunoassay using coated tubes. The same anti-HCG serum was employed in all immunoassays. When relatively crude HCG preparations (< 6000 IU/mg) were assayed, there was a highly significant (P < 0.001) correlation between the estimates obtained by all methods. However, when highly purified HCG preparations (> 6000 IU/mg) were assayed, the bioassay gave significantly higher potency estimates than the immunoassays. Furthermore, the estimates obtained by radioimmunoassays were significantly higher than those obtained by haemagglutination inhibition. However, the results obtained by the two radioimmunoassays showed a highly significant (P < 0.001) correlation. The regression coefficient of the line relating the logit transform of the response to loge, of the dose was significantly higher (b =−1.700±0.7322)) in assays employing the double antibody technique (a non-equilibrium system) than in solid phase (b =−0.945 ± 0.116) immunoassays (an equilibrium system). When highly purified HCG preparations were assayed by the double antibody technique, the regression coefficients were significantly higher than those obtained with the standard preparations. In relation to their biological activity, HMG preparations contained much less immunological activity, than HHG preparations. In addition, the regression lines obtained with HCG, HMG and HHG preparations showed significant deviations from parallelism when estimated by radioimmunoassays. The precision and sensitivity of the four assay methods increased in the following order: bioassay, haemagglutination inhibition, solid phase radioimmunoassay and radioimmunoassay involving the use of the double antibody technique. It is concluded that in relation to their biological activity, marked differences exist in the immunological activities of highly purified and relatively crude HCG preparations. In addition, the immunological activities of HCG, HMG and HHG preparations are dissimilar, when immunoassayed by the use of an anti-HCG serum.

MEASUREMENT OF RAT LH WITH A DOUBLE-ANTIBODY SOLID-PHASE RADIOIMMUNOASSAY: EFFECT OF LH-RH AND OF TESTOSTERONE OENANTHATE IN CASTRATED ANIMALS

1975 ◽  
Vol 78 (3) ◽  
pp. 451-460 ◽  
Author(s):  
J. S. E. Dericks-Tan ◽  
Hans-Dieter Taubert
Keyword(s):  
Solid Phase ◽  
Test System ◽  
Subsequent Treatment ◽  
Male Rats ◽  
Ventral Prostate ◽  
Phase Method ◽  
Double Antibody ◽  
Serum Lh ◽  
Solid Phase Radioimmunoassay ◽  
Lh Rh

ABSTRACT A double-antibody solid-phase radioimmunoassay for the measurement of rat LH was described. The use of a solid-phase method for the second incubation resulted in a considerable saving of time as compared to the conventional double-antibody method. The sensitivity and accuracy of the method were not affected by this modification. It was found that there was a cross-reaction between the NIAMD-anti-rat-LH-serum-1 used and NIAMD-rat-FSH-RP-1. This indicates that the test system does not specifically measure rat LH but gonadotrophic activity. The RIA was tested under different experimental conditions. In male rats, the effect of castration and of subsequent treatment with testosterone oenanthate upon serum LH, and upon the weight of the seminal vesicles, the ventral prostate, and the levator ani, was examined. Serum LH rose rapidly after castration and reached 2 to 4 weeks after orchidectomy a maximum (8–9 fold increase). Subsequent treatment with testosterone oenanthate caused a rapid decrease of serum LH. The accessory sex organs showed reciprocal changes of weight. In oophorectomized rats whose pituitaries had been blocked with oestradiol monobenzoate and progesterone, a linear log-dose response of plasma LH was found after administration of various doses of synthetic LH-RH.

Direct solid-phase radioimmunoassay for screening 17 alpha-hydroxyprogesterone in whole-blood samples from newborns.

1985 ◽  
Vol 31 (7) ◽  
pp. 1127-1130 ◽  
Author(s):  
L F Hofman ◽  
J E Klaniecki ◽  
E K Smith
Keyword(s):  
Filter Paper ◽  
Whole Blood ◽  
Solid Phase ◽  
Cross Reactivity ◽  
Blood Samples ◽  
Standard Curve ◽  
Logit Transformation ◽  
Analytical Recovery ◽  
Solid Phase Radioimmunoassay ◽  
Direct Solid

Abstract We describe a direct, solid-phase RIA for 17 alpha-hydroxyprogesterone (17-OH-P) that we are using to screen neonates for congenital adrenal hyperplasia. Phosphate buffer containing danazol and anti-17-OH-P is placed in tubes coated with antibody to IgG. The tubes also contain standards, controls, or blood samples on filter paper discs 3 mm in diameter. 125I-labeled 17-OH-P is added to each tube. The mixture is vortex-mixed and incubated overnight. The fluid and disc are removed, the radioactivity remaining in the tubes is counted, and the amount of 17-OH-P per disc is calculated by using a log-logit transformation of the standard curve. Results compare favorably with those by two extraction assays. Inter- and intra-assay CVs were less than 11% and less than 9%, respectively. Sensitivity was 2 pg per assay tube. There is no significant cross reactivity with structurally related steroids at their physiological concentrations. Analytical recovery of added 17-OH-P averaged 104%. 17-OH-P in whole blood spotted on filter paper is stable for at least six months.

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