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At least 19 recordsLinked to original sources
PubMed1991

Quantitation of HLA antigens on red cells by immunoradiometric assay.

Immunoradiometric assay was employed to quantitate HLA antigens on red cells. Using this technique HLA-B7, HLA-B17 and HLA-A28 were detected on the red cells of all individuals studied irrespective of the serological status of the Bennett-Goodspeed (Bg) antigens. However, HLA antigenic sites for serologically Bg positive red cells were significantly more than that for Bg negative red cells (P less than 0.001). Bga positive red cells possessed maximum number of antigenic sites as compared to Bgb and Bgc positive red cells.

Erythrocytes↗
PubMed1995

Measurement of urinary free beta-human chorionic gonadotropin by immunoradiometric assay.

An immunoradiometric assay for free beta-human chorionic gonadotropin (F beta hCG) is now available. Measurement of serum F beta hCG has been shown to be valuable for the diagnosis of trophoblastic disease and for screening Down's syndrome pregnancies. Urine specimens may be preferable to blood samples since collection of urine specimens usually are less inconvenient to patients than venipuncture. We have evaluated whether the immunoradiometric assay can be applicable to urine samples. The assay was sensitive and precise. When urine samples were diluted 4-fold or more, the diluted samples gave quantitative values, and recovery of beta hCG added to urine samples was satisfactory. Creatinine corrected urinary F beta hCG levels correlated with serum F beta hCG levels. Thus, the immunoradiometric assay was considered to be applicable to urine samples. Serum F beta hCG to hCG ratio has been reported to be important to distinguish among normal pregnancies, hydatidiform mole and choriocarcinoma. However, urinary F beta hCG to hCG ratio did not significantly correlate with serum F beta hCG to hCG ratio. And therefore, the clinical value of urinary F beta hCG to hCG ratio should be further investigated.

Chorionic Gonadotropin↗
PubMed1987

Measurement of corticotropin in unextracted plasma: comparison of a time-resolved immunofluorometric assay and an immunoradiometric assay, with use of the same monoclonal antibodies.

We describe a time-resolved immunofluorometric assay (IFMA) for corticotropin in unextracted human plasma, based on the use of two monoclonal antibodies: europium-labeled antibody 1A12 and antibody 2A3 coated onto microtiter wells. We compared the results of this assay with those of an immunoradiometric assay (IRMA) performed with the same antibodies working ranges (CV less than 10%) were 25 to 1000 ng/L and 22 to 1000 ng/L for the IFMA and IRMA, respectively, and both assays had comparable detection limits (IFMA 4.0 +/- 1 ng/L, IRMA 3.5 +/- 0.8 ng/L). Results by both assays for 130 patients' samples containing corticotropin within the range 3-100 ng/L and greater than ng/L correlated well (r = 0.88 and 0.92, respectively), and samples with corticotropin in the range 80-624 ng/L gave results that paralleled those for the standard curve. Corticotropin concentrations in apparently healthy subjects were consistent with those reported previously. The IFMA is a simple, precise, and robust assay that can be completed within one day. Its nonisotopic label is stable for at least 50 weeks.

Adrenocorticotropic Hormone↗
PubMed1993

Measurement of insulin-like growth factor-II in human plasma using a specific monoclonal antibody-based two-site immunoradiometric assay.

An immunoradiometric assay (IRMA) for the measurement of insulin-like growth factor-II (IGF-II) in human plasma has been developed, optimized and evaluated clinically in normal subjects and patients with disorders of the GH/IGF-I axis. Six monoclonal antibodies (MAbs) to recombinant human IGF-II (rhIGF-II) were produced, all of which had low cross-reactivity with rhIGF-I (< 0.01%) and insulin (< 0.01%). Compatibility of pairs of MAbs was tested in two-site IRMAs using three radioiodinated MAbs and three MAbs linked to Sephacryl S-300 (with separation of bound and free radiolabelled MAb by sucrose layering). Seven pairs of MAbs bound rhIGF-II and the combination of 125I-labelled W3D9 and W2H1 linked to solid phase was selected. The optimized assay had a completion time of 4 h, a minimum detection limit of 30 ng/ml (2.5 standard deviations from the zero standard) and detected a single peak of endogenous IGF-II in normal plasma which co-eluted with rhIGF-II after acid gel chromatography. IGF-II was measured in formic acid/acetone extracts of plasma from 16 normal subjects (mean 685, range 516-1008 micrograms/l), four acromegalic patients (mean 637, range 553-700 micrograms/l), fourteen patients with type-1 diabetes (mean 635, range 247-753 micrograms/l), nine patients with uraemia (mean 423, range 78-850 micrograms/l), and three patients with Laron-type GH insensitivity (75, 35 and 36 micrograms/l). No significant fluctuations were detected between samples obtained hourly from 08.00 to 19.00 h in normal subjects. Low levels of IGF-binding proteins (IGFBPs) remaining in plasma extracts may interfere with the measurement of IGF-II and give rise to falsely elevated IGF-II levels in radioimmunoassays or falsely suppressed levels in IRMAs. Such interference did not occur with the IRMA when used to measure IGF-II in extracts from normal subjects, acromegalic patients and patients with type-1 diabetes, and the addition of excess rhIGF-I in order to displace IGF-II from residual IGFBPs had no effect on IGF-II measurements in these samples. However, levels of IGF-II measured in extracts from patients with Laron-type GH insensitivity and patients with uraemia increased markedly after preincubation with excess rhIGF-I. The accurate measurement of IGF-II by IRMA in extracts from these subjects therefore requires the displacement of IGF-II from IGFBPs prior to assay. We conclude that, in contrast to radioimmunoassays, the two-site IRMA developed here provides a practical, rapid and specific method for the measurement of IGF-II in human plasma.

Antibodies, Monoclonal↗
PubMed1986

Interleukin 2 receptors are released by cells in vitro and in vivo. I. Detection of soluble IL 2 receptors in cell culture supernatants and in the serum of mice by an immunoradiometric assay.

An immunoradiometric assay (IRMA) detecting nanogram amounts of soluble mouse interleukin 2 receptors (IL 2R) is described. The IRMA is based on the finding that the two monoclonal antibodies AMT-13 and 7D4 react with two different epitopes of the mouse IL 2R molecule. As detected by IRMA, T lymphoblasts and IL 2R+ leukemic cells when incubated release IL 2R in soluble form into the culture medium. In the serum of mice inoculated with syngeneic IL 2R+ leukemic cells, growing as a solid tumor, IL 2R were detectable. The results raise the question whether circulating IL 2R are involved in down-regulation of immune responses.

Animals↗
PubMed1983

Detection of factor IX inhibitors by immunoradiometric assay.

An immunoradiometric assay (IRMA) for the determination of factor IX inhibitors in haemophilia B was developed. The assay was based on competitive binding between radiolabelled anti-IX and inhibitors in the test material of immobilized IX:C. 5 haemophiliacs with known inhibitors were investigated with the new method and with a conventional neutralization test. An inhibitor titre as low as 5 X 10(-4) U/ml could be measured by IRMA, showing it to be about 500 times as sensitive as the conventional neutralization assay used, and in 2 cases the inhibitors could only be detected by IRMA. The new method is thus useful for the investigation of patients with low inhibitor titres, or when inhibitors with divergent properties are suspected.

Antibodies↗
PubMed1988

Direct measurement of the precursors of adrenocorticotropin in human plasma by two-site immunoradiometric assay.

An immunoradiometric assay (IRMA) for the direct measurement of the precursors of ACTH in unextracted human plasma has been developed and evaluated clinically in normal subjects and patients with disorders of the hypothalamic-pituitary-adrenal axis. The IRMA is based on an iodinated monoclonal antibody to ACTH and a monoclonal antibody to gamma MSH coupled to Sephacryl S300. The assay detects only peptides containing both epitopes, i.e. POMC (31K) and pro-ACTH (22K). The reference standard was partially purified POMC from culture medium of human corticotroph adenoma cells. The detection limit (greater than +2.5SD of the 0 standard) was 2.0 pmol/L and the within-assay coefficient of variation was less than 10% between 29 and 2600 pmol/L. Plasma concentrations of ACTH precursor peptides in 11 normal subjects sampled at 0930 h ranged from 5-34 pmol/L. The concentrations in the patient groups studied were: 260-2300 pmol/L in 5 patients with the ectopic ACTH syndrome associated with small cell lung cancer, less than 2.0-104 pmol/L in 10 patients with pituitary-dependent Cushing's disease, 23 pmol/L in a patient with Nelson's syndrome, and 3.0-230 pmol/L in 5 patients with Addison's disease. We conclude that this IRMA offers a simple and reliable method for measuring ACTH precursors in unextracted plasma. The proportionately greater elevation of ACTH precursors compared to ACTH in patients with the ectopic ACTH syndrome associated with small cell lung cancer but not in pituitary-dependent Cushing's syndrome, suggests that this assay may be clinically useful.

ACTH Syndrome, Ectopic↗
PubMed1989

Influence of matrix on concentrations of somatotropin measured in serum with commercial immunoradiometric assays.

Using immunoradiometric assays (IRMAs) from Hybritech Inc. (H) and Nichols Institute Diagnostics (ND), we measured somatotropin (human growth hormone, hGH) in serum samples obtained every 20 min for 24 h from 10 prepubertal subjects with short stature. Results obtained with the ND reagents were 2.74 times greater than those obtained with the H reagents (P = 0.00001, r = 0.94, SEE = 3.9, n = 720). We therefore compared the IRMAs with the standard hGH RIA from the National Institutes of Health (NIH) National Hormone and Pituitary Program, using the genetically engineered hGH preparations (from Genentech Inc.) 22-kDa hGH and methionated 20-kDa hGH. We also assayed human pituitary hGH (NIH, lot no. AFP-4793B). Each hGH preparation was diluted in three diluent buffer systems: horse serum from H and from ND, and human serum. The RIA and H-IRMA gave superimposable standard curves for all hGH preparations in each diluent. The methionated 20-kDa hGH was not detected in the H-assay. Use of human serum matrix in the ND-IRMA shifted the standard curve as compared with the horse-serum matrix, giving equivalent binding at lower concentrations; i.e., serum hGH was overestimated in samples assayed against standards diluted in horse serum. Quality-control materials (Ciba-Corning) yielded disparate results in all three assays, yet human serum pools containing hGH gave similar results in the H and the NIH assays, and higher values in ND. When a human serum standard was used in the ND assay, both IRMAs gave similar results to the RIA assay for human serum samples. Reference intervals for hGH should be determined by each analytical laboratory, to prevent misdiagnosis of patients. Furthermore, quality-control material should be of human origin, because commercially supplied quality-control material does not react the same as human serum in some hGH assays.

Child↗
PubMed1985

Thyrotropin as measured by a sensitive immunoradiometric assay.

An immunoradiometric assay (Boots-Celltech's "Sucrosep"; IRMA) for thyrotropin (TSH) was evaluated and results were compared with those of our in-house RIA procedure. The IRMA had a sensitivity of 0.02 milli-int. unit/L. In addition it displayed excellent intra- and inter-batch precision, cross-reacted negligibly with other pituitary hormones, and appeared to be relatively free of matrix effects (although deionized water did give an apparent TSH concentration of 0.13 milli-int. unit/L). Over the range 1.2 to 64 milli-int. units/L, correlation between the IRMA (y) and RIA (x) was excellent: y = 1.16x - 1.69 (r = 0.98). The normal reference interval for the IRMA was 0.4 to 4.2 milli-int. units/L. For patients suspected of hyperthyroidism who had been subjected to the thyroliberin test, the IRMA more effectively differentiated between euthyroidism and hyperthyroidism. During testing with thyroliberin, all nonresponding hyperthyroid patients had initial baseline TSH concentrations of less than 0.02 milli-int. unit/L by IRMA. This sensitive assay represents an important advance in the ability to differentiate between euthyroidism and hyperthyroidism.

Humans↗
PubMed1995

No evidence of the hook effect in peritoneal fluid CA 125 measurement using immunoenzymatic second generation assays: comparison with immunoradiometric assays.

OBJECTIVE: To monitor the occurrence of the hook effect in measurements of peritoneal fluid CA 125 levels using two different immunoenzymatic second generation assays (ETI-II and EIA-II), and to compare these results with those obtained using the respective immunoradiometric versions of the assays (IRMA-II and ELSA-II). STUDY DESIGN: CA 125 levels were determined in peritoneal fluid and serum samples obtained from 45 women with gynecological diseases. The assays were carried out using IRMA-II and ETI-II (Sorin Biomedica) and ELSA-II and EIA-II (CIS Bio International) assays. Occurrence of the hook effect and linearity of the assays were evaluated. Statistical analyses were performed by Wilcoxon's test and linear regression analysis. RESULTS: Undiluted peritoneal fluids, assayed for their CA 125 content, showed falsely underestimated values of the antigen when IRMA-II and ELSA-II assays were performed. The phenomenon disappeared only at high dilutions of the sample (> 50). Conversely, immunoenzymatic ETI-II and EIA-II assays performed on undiluted peritoneal fluids did not show underestimated CA 125 values. CA 125 values obtained by immunoenzymatic assay were lower than those obtained using their respective immunoradiometric versions at a dilution of 1:100 (P < 0.001). A good correlation was observed between ELSA-II and EIA-II (r = 0.929) CA 125 values. CONCLUSION: The EIA-II immunoenzymatic assay appears to be more suitable for CA 125 measurement in peritoneal fluid in that it is not subject to the hook effect and its results correlated well with those obtained via its immunoradiometric version.

Ascitic Fluid↗
PubMed1992

Glycosylated serum protein levels assayed with highly sensitive immunoradiometric assay accurately reflect glycemic control of diabetic patients.

OBJECTIVE: To develop a sensitive and reliable immunoreadiometric assay to measure glycosylated lysine residues on serum proteins (GSP) and to evaluate its efficacy in monitoring glycemic control. RESEARCH DESIGN: The effect of acute and chronic in vitro and in vivo changes in glucose levels on GSP concentration was evaluated. GSP determinations from insulin-dependent diabetic (IDDM) patients, non-insulin-dependent diabetic (NIDDM) patients, and control subjects were correlated with other indices of glycemic control. RESULTS: The GSP levels were unaffected by acute glucose changes after food or intravenous glucose administration but increased during storage at -20 degrees C due to in vitro glycosylation by endogenous glucose. Immediate acidification of the serum prevented this, permitting long-term storage despite high ambient glucose levels. In randomly selected diabetic patients, 96% of GSP values were greater than the mean +3SD of nondiabetic control subjects. In diabetic patients, GSP levels correlated with mean plasma glucose concentrations (Kendall correlation statistics 0.47, P less than 0.001), fasting plasma glucose levels (Kendall statistics 0.42, P less than 0.001), and glycosylated hemoglobin (GHb, Kendall statistics 0.30, P less than 0.005). Induction of near-normal glycemia in poorly controlled NIDDM patients reduced GSP levels with a slope consistent with a half time of disappearance of 4.7 +/- 0.4 days. GSP levels remained elevated in 6 of 10 well-controlled NIDDM patients, despite normal GHb concentrations. Chronic hypoglycemic states, like pregnancy and hyperinsulinemic hypoglycemia, were associated with significantly low GSP levels. CONCLUSION: We describe a reproducible and sensitive immunoradiometric assay for GSP that closely reflects the degree of glycemic control in diabetic patients. Further studies are needed to determine whether this assay may be useful in screening for glucose intolerance or gestational diabetes.

Blood Glucose↗
PubMed2006

Chromogranin A immunoradiometric assay in diagnosis of pheochromocytoma: comparison with plasma metanephrines and 123I-MIBG scan.

AIM: The aim of this study is to investigate the diagnostic performance of serum chromogranin A (CgA) and plasma metanephrines (MN) assays in the diagnosis of pheochromocytoma. METHODS: We enrolled 44 patients affected by histologically proved adrenal pheochromocytoma. All patients underwent abdominal computed tomography and whole body 123I-MIBG scan to stage the disease. One hundred healthy blood donors and 148 patients affected by essential hypertension were enrolled as controls. Serum CgA and plasma MN were assayed by immunoradiometric assay (IRMA) and high-performance liquid chromatography. Cut-off levels were selected to obtain 99% specificity in healthy control subjects. RESULTS: Both MN and CgA showed 95% sensitivity with comparable high specificity and diagnostic accuracy (96% and 96% for CgA, 94% and 95% for MN, respectively). By employing both markers and considering CgA or MN positivity, a 100% sensitivity was obtained with 95% accuracy. MN and CgA concentration clearly increased from controls to patients with pheochromocytomas (P<0.0001). A relationship between serum CgA (but not MN) and tumor mass was found (P<0.0001). CONCLUSIONS: Both markers are sensitive and specific in chromaffin-tumors detection: CgA IRMA assay employed a simple and feasible technology and showed to be as sensitive and slightly more accurate than MN.

3-Iodobenzylguanidine↗
PubMedDate not supplied

Evaluation of the IMMULITE BR-MA and CEA assays and comparison with immunoradiometric assays for CA15-3 and CEA in breast cancer.

This study was designed to evaluate the performance of a new automated assay system, the IMMULITE Automated Immunoassay Analyzer in comparison with more commonly used IRMA assays for measuring circulating tumour marker levels in breast cancer patients. The assays investigated measure the MUC1 mucin (CA15-3 antigen) or CEA. Serum samples from breast cancer patients with various stages of disease and from a group of normal individuals were analysed. Significant correlations were found between tumour marker levels measured using the IMMULITE BR-MA and the CA15-3 assays and between levels measured using the two CEA assay formats. Levels of IMMULITE BR-MA and CEA correlated with stage of disease suggesting that both are markers of tumour burden Levels in Stage III breast cancer patients were found to be significantly higher than those of normals using the IMMULITE system but not the IRMA assays. This would suggest a role for the automated system in the monitoring of breast cancer at an earlier stage than that at which tumour markers are routinely used. Elevated marker levels did not correspond to any particular site of metastasis however, a greater proportion of patients with multiple sites of metastasis had elevated levels compared with those with secondary disease at a single site. We conclude that the IMMULITE Automated Immunoassay Analyzer is of value in the routine surveillance of tumour marker levels.

Automation↗
PubMed1981

Immunoradiometric assay of human leukocyte interferon using monoclonal antibody.

Interferon research has been hampered by problems in its assay. The only widespread assays use tissue culture cells and compare some parameter of viral growth (such as viral RNA synthesis or host cell death) in the presence and absence of interferon. These complex biological assays, though sensitive, are laborious and subject to inherent variability. In particular, components other than interferon present in the assay sample often influence viral growth. There is clearly a need for a simple, direct, interferon assay; I now describe such an assay--an immunoradiometric assay utilizing monoclonal antibody.

Antibodies↗
PubMedDate not supplied

Properties, variants, and applications of the immunoradiometric assay method.

The immunoradiometric assay (IRMA) and 2-site IRMA techniques employ purified radioactive antibodies to convert soluble antigens into a directly detectable complex. Unreacted labelled antibody is discarded by reaction with solid-phase antigen (IRMA), or by a preliminary insolubilization of the unknown antigen using solid-phase antibody (2-site IRMA). The preparation and properties of antigen-immunoadsorbent, solid-phase antibody, and purified radioactive antibodies are discussed. Analysis of the dose-response curve and problems in dose-interpolation are similar in IRMA and 2-site IRMA, and different from radioimmunoassay and other competitive-binding assay systems. The most significant assay variables include: exchange of radioactive antibody in IRMA reaction 2, stability and reactivity of the solid-phase antibody, washing the solid-phase, non-specific interference by serum proteins, and a paradoxical fall in tube radioactivity at high dose (2-site IRMA) (the 'high-dose hook effect'). 2-site IRMA has advantages in specificity (by using separate antibodies directed to 2 different antigen sites), sensitivity and assay range (because of the low zero dose-response). Assay variants include: alternative antibody labels, various solid-phase antibody preparations (including immunological 'spacer-arms'), non-immunological insolubilization of antigen, univalent radioactive antibodies, repeated immunological extractions of unknown antigen, and non-immunological separations in IRMA reaction 2. Both IRMA and 2-site IRMA have been adapted to the use of labelled anti-IgG as an additional 'universal' reagent, thereby avoiding the necessity for the preparation of radioactive antibodies specific for the unknown antigen. These techniques have been applied to the assay of a great variety of antigens and will be especially appropriate for automation.

Antibodies↗
PubMed1980

Serum ferritin concentration and isoferrtin patterns detected by immunoradiometric assay.

Two-site immunoradiometric assay for serum ferritin was developed by anti-human liver ferritin antiserum applied on polyvinyl V-bottom microtiter plates. The coefficient of variation was 4.4-5.9% of liver ferritin dissolved in 1/4 diluted human serum at a concentration of 0.625-125 ng/ml. The lowest concentration (0.625 ng/ml) was statistically distinguishable from the background by Student's t test (p < 0.05). Isoferritin patterns of serum ferritin and tissue ferritins were examined by the combination of gel isoelectric focussing and 2-site immunoradiometric assay in order to explore any similarities in pIs within these ferritins. Although serum ferritin had wide range of pIs, its basic isomers corresponded well to those of liver and spleen ferritin. Geometric means and their 95% confidence limits in serum ferritin concentration were 70 ng/ml and 12-411 ng/ml, respectively, in sera of 205 healthy males (16-25 years), and 12 ng/ml and 1-211 ng/ml, respectively, in 421 healthy females (16-59 years).

Adolescent↗
PubMed1987

Effect of nonspecific interactions of the hepatitis B surface antigen with the solid phase on its detection by two-site immunoradiometric assay.

Two-site immunoradiometric assay (Austria II-125, procedure B, Abbott) was used to detect hepatitis B surface antigen (HBsAg) in chronic carrier sera from clinically healthy subjects or from kidney transplant or hemodialysis patients. The titration curves of some high-titered sera showed a prozone-like effect; this corresponded in two such sera LB and MA, to congruent to 80% inhibition of HBsAg detection in undiluted serum relative to a 1:800 dilution. The nature of the inhibition in the above two sera was investigated. We found that the inhibition depended on the time of first incubation but not on the affinity of capture antibodies. Nonspecific adsorption of HBsAg to the solid phase during the first incubation with inhibitory sera and elution of the thus adsorbed antigen during the second incubation, with the resulting neutralization of 125I-labeled anti-HBs, appeared to cause the inhibition of detection observed. The inhibition was prevented by addition of the detergent Triton X-100 to serum or by elution of adsorbed antigen prior to the second incubation, thus indicating that further improvements of the HBsAg immunoradiometric assay may be possible.

Adsorption↗
PubMed1984

Optimization of assay conditions in monoclonal-based immunoradiometric assay.

The production of relatively large amounts of pure monoclonal antibodies (MAb) has facilitated the development of MAb-based immunometric assays for a variety of clinically important analytes. 'Two-site' heterogeneous assays are now available which incorporate a labelled MAb and a second, different MAb coupled to a solid support. These assays possess certain advantages over the corresponding immunoassays including speed, precision, working range and specificity. They are largely dependent on two specific molecular interactions (labelled MAb-antigen; antigen--solid support MAb) and thus one might expect them to be particularly sensitive to assay conditions. With reference to two MAb--based two-site immunoradiometric assays (for human growth hormone and prolactin) which are being developed in this laboratory we wish to report the effect of various conditions including pH, ionic strength, buffer species etc. on assay response in order to emphasize the need for careful optimisation of monoclonal antibody based assays.

Animals↗