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.

Solid-phase radioimmunoassay for morphine, with use of an affinity-purified morphine antibody.

1978 ◽  
Vol 24 (2) ◽  
pp. 339-342 ◽  
Author(s):  
M Steiner ◽  
J L Spratt
Keyword(s):  
Affinity Chromatography ◽  
Correlation Coefficient ◽  
Solid Phase ◽  
Rapid Determination ◽  
Biological Fluids ◽  
Inhibitory Effect ◽  
Major Metabolite ◽  
Solid Phase Radioimmunoassay ◽  
Related Compounds

Abstract Morphine antibody purified by affinity chromatography was used to develop a solid-phase radioimmunoassay for morphine in polystyrene tubes. The tubes are coated with an appropriate concentration of the purified antibody, rinsed three times with buffered saline, and stored at -15 degrees C. Using tritiated dihydromorphine, we determined competitive morphine binding by difference when the radioactivity in the assay supernates was measured after incubation (1 h, 37 degrees C). Five standard curves, with use of serum equivalents of morphine ranging from 0 to 6 mug/liter, were linear and had a mean correlation coefficient of 0.98. Uncer conditions of the assay, levorphanol was comparable to morphine in its inhibitory effect on binding of labeled dihydromorphine, whereas dextrorphan was essentially inactive. Morphine-3-glucuronide, a major metabolite, is 55-fold less inhibitory in terms of its capacity to displace the radiolabel. We believe that the sensitivity of the technique, coupled with the simplicity of nonseparatory sampling, renders the system suitable for rapid determination of morphine and related compounds in biological fluids.

Direct chemiluminescence immunoassay of estradiol in saliva

1990 ◽  
Vol 36 (12) ◽  
pp. 2036-2041 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
J Bouve ◽  
D Leyseele ◽  
D VandeKerckhove
Keyword(s):  
Detection Limit ◽  
Incubation Time ◽  
Solid Phase ◽  
Chemiluminescence Immunoassay ◽  
Microtiter Plates ◽  
Analytical Recovery ◽  
Plate Reader ◽  
Steroid Binding ◽  
Method R ◽  
Direct Solid

Abstract A sensitive and simple direct solid-phase chemiluminescence immunoassay is described for estradiol in saliva. In this assay, a second antibody is bound to the wells of microtiter plates. Either buffer with standards or saliva (100 microL) is incubated in these wells with monoclonal anti-estradiol antibody and with estradiol-isoluminol conjugate. Incubation time is 2 h. Chemiluminescence of the bound fraction is measured in a manually operated luminometer (Biocounter). The assay has a detection limit of 3.8 pmol/L; analytical recovery of added estradiol is 96.8% (SD 7.0%); within- and between-assay CVs range between 2.5% and 12.7%. Forty unknown saliva samples can be assayed and results calculated within 4.5 h. Results of a slightly modified procedure-with black microtiter plates and a prototype of an automated plate reader (Luminoskan)--compare well with those of the described method (r = 0.97). Because steroid-binding globulins have been found in saliva, the effect of displacing agents on the results of the direct chemiluminescence assay is described.

Evaluation of a direct solid-phase radioimmunoassay for progesterone, useful for monitoring luteal function.

1984 ◽  
Vol 30 (2) ◽  
pp. 284-286 ◽  
Author(s):  
N P Kubasik ◽  
G D Hallauer ◽  
R G Brodows
Keyword(s):  
Detection Limit ◽  
Luteal Phase ◽  
Solid Phase ◽  
Cross Reactivity ◽  
Luteal Function ◽  
Menstrual Cycles ◽  
Analytical Recovery ◽  
Solid Phase Radioimmunoassay ◽  
Clinically Significant ◽  
Direct Solid

Abstract We have evaluated a commercially available, direct, solid-phase radioimmunoassay kit for progesterone determination in serum or plasma. The assay is precise, within-run precision (CV) in the clinically significant ranges being 2.5 to 5.2%, between-run 5.5 to 5.8%. Mean analytical recovery of different concentrations of progesterone added to serum was 99.7% (range 95.3 to 102.7%). Fourteen closely related steroids showed no cross reactivity. The minimum detection limit was 0.5 microgram/L. Luteal-phase progesterone concentrations in serum were increased (greater than 3 micrograms/L) in 19 normal ovulatory menstrual cycles and decreased (less than 1.5 micrograms/L) in two nonovulatory cycles. We found this direct assay for progesterone to be analytically and clinically sound, and useful for assessing luteal-phase function.

Solid-phase radioimmunoassay for human placental lactogen in serum.

1983 ◽  
Vol 29 (1) ◽  
pp. 168-170 ◽  
Author(s):  
G Samuel ◽  
N Sivaprasad ◽  
K B Shah ◽  
R S Mani
Keyword(s):  
Ionic Strength ◽  
Incubation Time ◽  
Solid Phase ◽  
Placental Lactogen ◽  
Human Placental Lactogen ◽  
Optimum Conditions ◽  
Nonspecific Adsorption ◽  
Assay Tube ◽  
Solid Phase Radioimmunoassay ◽  
Assay Technique

Abstract We report a solid-phase radioimmunoassay for human placental lactogen, in which antibody-coated polystyrene tubes and polyurethane sponges are used. Incubation time and temperature, ionic strength of buffer, and nonspecific adsorption have been evaluated for optimum conditions required for routine clinical assays. The validity of the assay technique has been ascertained by its specificity, reproducibility, and recovery. The sensitivity of this method is 0.4 ng of placental lactogen per assay tube.

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."

An "antibody electrode," preliminary report on a new approach in enzyme immunoassay.

1979 ◽  
Vol 25 (2) ◽  
pp. 318-321 ◽  
Author(s):  
J L Boitieux ◽  
G Desmet ◽  
D Thomas
Keyword(s):  
Enzyme Immunoassay ◽  
Surface Antigen ◽  
Solid Phase ◽  
Biological Fluids ◽  
Hepatitis B Surface Antigen ◽  
Antigen Concentration ◽  
New Approach ◽  
Solid Phase Radioimmunoassay ◽  
Reliable Procedure ◽  
Sensitive Electrode

Abstract We report preliminary experiments on a new, sensitive, and reliable procedure for enzyme immunoassay of various antigens in biological fluids. The method, developed from the biological model "hepatitis B surface-antigen/antibodies," is less time consuming than most immunochemical techniques and eliminates many inconveniences arising from use of isotopes. We use a solid-phase "sandwich" procedure, the antibodies being immobilized on gelatin membranes, and determine antigen concentration with the help of an iodide-sensitive electrode modified by fixing the active membrane onto the crystal sensor. We have established the analytical criteria of the method and compared it with the solid-phase radioimmunoassay for the surface antigen in dilution series. One-tenth microgram of antigen per liter can be reproducibly detected with our method. Use of antibody electrode should easily be extended to assay of other antigens and haptens that usually are determined by radioimmunoassay.

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.

SOLID-PHASE RADIOIMMUNOASSAY OF ESTROGENS IN BIOLOGICAL FLUIDS

1970 ◽  
Vol 65 (1_Suppl) ◽  
pp. S332-S346 ◽  
Author(s):  
G. E. Abraham ◽  
W. D. Odell ◽  
R. Edwards ◽  
J. M. Purdy
Keyword(s):  
Detection Method ◽  
Solid Phase ◽  
Biological Fluid ◽  
Direct Method ◽  
Biological Fluids ◽  
Ether Extract ◽  
Human Follicular Fluid ◽  
Male And Female ◽  
17Β Estradiol ◽  
Solid Phase Radioimmunoassay

ABSTRACT Using antibody-coated test tubes as a detection method, three types of assay for 17β-estradiol (E2) have been investigated. Method A involved the measurement of E2 in the dried ether extract of biological fluids. Method B required a chromatographic step after ether extract, prior to measurement of E2. Method C (direct method) measured E2 directly in the biological fluid, diluted with assay buffer. Method C was found reliable for the measurement of E2 in human follicular fluid, but not in pregnancy serum. Method A was adequate for pregnancy serum, but overestimated E2 in serum from normal adult male and female subjects. Method B was reliable to measure E2 in all the biological fluids tested.

Radioimmunoassay of Fibrinogen and Its Proteolysis Products

1968 ◽  
Vol 20 (01/02) ◽  
pp. 001-006 ◽  
Author(s):  
K. J Catt ◽  
J Hirsh ◽  
D. J Castelan ◽  
H. D Niall ◽  
G. W Tregear
Keyword(s):  
Solid Phase ◽  
Solid Phase Radioimmunoassay ◽  
Radioimmunoassay Method

SummaryThe solid-phase radioimmunoassay method has been applied to the measurement of fibrinogen. The method is extremely sensitive, being able to detect fibrinogen concentrations as low as 10 ng/ml. The immunoreactivity of fibrinogen proteolysis products differs from that of native fibrinogen, early proteolysis products showing enhanced immunoreactivity which decreases progressively with further digestion.

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