Radioreceptor Assay for Quantifying FK-506 Immunosuppressant in Whole Blood

1992 ◽  
Vol 38 (7) ◽  
pp. 1307-1310 ◽  
Author(s):  
J N Murthy ◽  
Y Chen ◽  
V S Warty ◽  
R Venkataramanan ◽  
J G Donnelly ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Whole Blood ◽  
Binding Protein ◽  
Radioreceptor Assay ◽  
Fk 506 ◽  
Blood Concentrations ◽  
Chromatographic Columns ◽  
Analytical Recovery

Abstract We describe a quantitative radioreceptor assay (RRA) for quantifying FK-506 in whole blood. FK-506 extracted from whole blood with a cyclohexyl-sorbent column competes with [3H]dihydro-FK-506 for binding to a partially purified preparation of FK-506 binding protein (FK-BP). Free and protein-bound FK-506 are separated on LH 20 Sephadex chromatographic columns. We compared the results of this method with those of an enzyme immunoassay that uses a monoclonal antibody: r = 0.97, Sy/x = 0.039. Between-day precisions (CV) at FK-506 concentrations of 8 and 17 micrograms/L were 9.2% and 8.2%, respectively. Within-run precisions were 5.9%, 8.1%, and 9.4%, respectively, at 4, 8, and 15 micrograms/L. Analytical recovery, evaluated at 5, 10, 15, 20, and 25 micrograms/L for FK-506 added to whole blood, ranged from 98% to 103%. The assay can reliably quantify FK-506 blood concentrations between 1.0 and 25 micrograms/L.

Plasma vs whole blood for therapeutic drug monitoring of patients receiving FK 506 for immunosuppression

1994 ◽  
Vol 40 (12) ◽  
pp. 2247-2253 ◽  
Author(s):  
M Winkler ◽  
B Ringe ◽  
J Baumann ◽  
M Loss ◽  
K Wonigeit ◽  
...  
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Enzyme Immunoassay ◽  
Drug Monitoring ◽  
Whole Blood ◽  
Plasma Concentrations ◽  
Therapeutic Monitoring ◽  
Therapeutic Drug ◽  
Fk 506 ◽  
Blood Samples ◽  
The Matrix

Abstract By retrospective analysis of 13,000 blood samples obtained from 248 patients receiving FK 506 therapy, we compared the suitability of plasma with that of whole blood as the matrix for therapeutic drug monitoring of FK 506. The plasma concentrations did not correlate with the concentrations in whole blood (r = 0.56). In contrast to plasma samples (analyzed by enzyme immunoassay), FK 506 was detectable in all whole-blood samples (analyzed by enzyme immunoassay/microparticle enzyme immunoassay). The inter- and intraindividual variations of FK 506 measurements were greater in plasma than in whole blood. Moreover, plasma concentrations correlated only poorly with clinical events. There was a tendency to greater plasma concentrations being measured during episodes of toxicity, but no clear difference was evident between stable course and rejection. In whole-blood specimens, a correlation between reduced or increased FK 506 concentrations and rejection or toxicity, respectively, was observed. The discriminatory power of whole-blood values was greater for the differentiation between toxicity and stable course than between rejection and stable course. We therefore recommend whole blood rather than plasma as the matrix for therapeutic monitoring of FK 506 concentrations.

Particle-enhanced turbidimetric inhibition immunoassay for theophylline evaluated with the Du Pont aca.

1984 ◽  
Vol 30 (11) ◽  
pp. 1870-1874 ◽  
Author(s):  
K E Opheim ◽  
M R Glick ◽  
C N Ou ◽  
K W Ryder ◽  
L C Hood ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Enzyme Immunoassay ◽  
High Performance ◽  
Correlation Coefficients ◽  
Comparison Of Results ◽  
Analytical Recovery

Abstract We evaluated the Du Pont Particle-Enhanced Turbidimetric Inhibition Immunoassay (PETINIA) for theophylline. The imprecision (CV) of the assay was less than 4.7% between-run and less than 3.6% within-run for theophylline concentrations between 5 and 30 mg/L. Standard curves for the assay were linear for theophylline concentrations from 0 to 46 mg/L and were stable throughout the study (i.e., for at least three months). The monoclonal antibody against theophylline used in this assay increases specificity; of the possibly interfering drugs, metabolites, and anticoagulants tested, only 1,3-dimethyluric acid and EDTA showed measurable effects. Bilirubin (less than 300 mg/L), hemoglobin (less than 6 g/L), or lipemia (triglycerides less than 6 g/L) does affect the quality of the assay. Analytical recovery of theophylline added to serum (5 to 40 mg/L) averaged 98% (range 93% to 112%). Comparison of results for patients' sera by the PETINIA method with those by enzyme immunoassay (EMIT) and by "high-performance" liquid chromatography yielded slopes and intercepts not significantly different from 1.0 and 0.0, respectively, and correlation coefficients ranging from 0.986 to 0.995.

Abbott TDx monoclonal antibody assay evaluated for measuring cyclosporine in whole blood

1990 ◽  
Vol 36 (11) ◽  
pp. 1969-1973 ◽  
Author(s):  
R W Yatscoff ◽  
K R Copeland ◽  
C J Faraci
Keyword(s):  
Monoclonal Antibody ◽  
Renal Transplant ◽  
Whole Blood ◽  
Cross Reactivity ◽  
The Other ◽  
Renal Transplant Recipients ◽  
Transplant Recipients ◽  
Antibody Assay ◽  
Analytical Recovery ◽  
Blood Specimens

Abstract We report here the evaluation of the Abbott TDx assay with a monoclonal antibody for selectively quantifying cyclosporine (CsA) in whole blood. Over the clinically relevant concentration ranges, results with this assay demonstrated within- and between-run CVs of less than 2.5% and 5%, respectively; sensitivity of 25 micrograms/L; good analytical recovery (100.3%); and linearity with whole-blood specimens. The percentage cross-reactivity of the major CsA metabolites varied from 15.3% for AM9 (M-1), 8.2% for AM1 (M-17), and 3.7% for AM4N (M-21), to less than 3% for the other metabolites tested. Results with the TDx assay (y) correlated well with those by the Sandimmune selective RIA (x; Sandoz) with blood specimens from 44 renal-transplant recipients (n = 44, x= 187.3, y = 198.9, y = 5.49 + 1.03x, r = 0.987). The TDx values were on average 24% higher than those by HPLC (x') with the same patients' specimens (n = 44, x' = 159.9, y = 198.9, y = 15.9 + 1.14x', r = 0.967). We conclude that the Abbott TDx monoclonal antibody assay provides a rapid, precise, and accurate means for quantifying CsA in whole blood.

Hypoxemia regulates effect of lipopolysaccharide on polymorphonuclear leukocyte CD11b/CD18 expression

1994 ◽  
Vol 76 (4) ◽  
pp. 1657-1663 ◽  
Author(s):  
H. H. Simms ◽  
R. D′Amico
Keyword(s):  
Monoclonal Antibody ◽  
Polymorphonuclear Leukocyte ◽  
Whole Blood ◽  
Lipid A ◽  
Cytochalasin B ◽  
Actinomycin D ◽  
H2o2 Production ◽  
Functional Capability ◽  
Receptor Shedding ◽  
Do So

Expression of the receptor CD11b/CD18 on the polymorphonuclear leukocyte (PMN) surface is important for several aspects of PMN function during endotoxemia. The mechanisms underlying regulation of CD11b/CD18 expression during hypoxemia and endotoxemia, however, are less clear. We investigated the effects of exposure of whole blood PMNs to lipopolysaccharide (LPS) during hypoxemia. During hypoxemia (10–30% O2 saturation), LPS reduced CD11b/CD18 expression. Both kinetic assays and experiments with microfilament stabilizers (phalloidin, cytochalasin B) demonstrated that this was most likely due to receptor shedding. Incubation of whole blood PMNs with an anti-CD14 monoclonal antibody (MEM18) largely prevented the LPS-induced reduction of CD11b/CD18 expression. Decreased CD11b/CD18 expression reduced PMN functional capability, as the binding of its ligand (erythrocytes opsonized with the 3rd component of complement Cbi) and intracellular H2O2 production were reduced. After exposure to LPS, N-formyl-methionyl-leucyl-phenylalanine could rapidly induce new CD11b/CD18 receptors to the cell surface, and this was not inhibited by actinomycin D or cycloheximide. After reoxygenation (> 90% O2 saturation), CD11b/CD18 expression was restored, and this was abrogated by exposure to cytochalasin B. Lipid A was able to reproduce the effects of LPS during hypoxemia and hypoxemia-reoxygenation but required a 10-fold greater concentration to do so. These results demonstrate that during hypoxemia an important pathophysiological property of LPS is to reduce CD11b/CD18 expression. This results in diminished PMN functional capability, which would contribute to the pathogenicity of LPS during acute infectious states.

Development of a Monoclonal Antibody-Based Enzyme Immunoassay for the Pyrethroid Insecticide Deltamethrin

2010 ◽  
Vol 58 (14) ◽  
pp. 8189-8195 ◽  
Author(s):  
Ye Kong ◽  
Qi Zhang ◽  
Wen Zhang ◽  
Shirley J. Gee ◽  
Peiwu Li
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Pyrethroid Insecticide

scholarly journals Bound/free separation methods in steroid enzyme immunoassay with monoclonal antibody.

1988 ◽  
Vol 36 (9) ◽  
pp. 3525-3531 ◽  
Author(s):  
HIROSHI HOSODA ◽  
REIKO TSUKAMOTO ◽  
SAKIKO TAMURA ◽  
TOSHIO NAMBARA
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay
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