Establishment of a Particle Concentration Fluorescence Immunoassay (PCFIA) for the Measurement of Human IgG in Culture Supernatants

1988 ◽  
pp. 85-100
Author(s):  
M. B. Compas ◽  
Y. G. Alevy
Keyword(s):  
Particle Concentration ◽  
Fluorescence Immunoassay ◽  
Human Igg ◽  
Culture Supernatants

Development and application of monoclonal antibodies to human cardiac myoglobin in a rapid fluorescence immunoassay

1991 ◽  
Vol 37 (8) ◽  
pp. 1356-1364 ◽  
Author(s):  
D P Silva ◽  
Y Landt ◽  
S E Porter ◽  
J H Ladenson
Keyword(s):  
Monoclonal Antibodies ◽  
Particle Concentration ◽  
High Dose ◽  
Fluorescence Immunoassay ◽  
Hook Effect ◽  
One Step ◽  
Analytical Importance ◽  
Rapid Kinetics ◽  
Kinetics Of Reaction ◽  
Kinetics Of

Abstract Myoglobin (Mb) is considered a useful marker for early detection of myocardial infarction and for monitoring cardiac reperfusion after thrombolytic therapy. We developed eight monoclonal antibodies to human cardiac Mb, characterized their epitopic reactivity, and determined which combinations of the antibodies are useful in two-site immunoassays. We configured two of the monoclonal antibodies in a one-step, two-site particle concentration fluorescence immunoassay (PCFIA) for measurement of Mb. The PCFIA has rapid kinetics of reaction, being complete in 15 min, and has a linear analytical range of 20-675 micrograms/L for human Mb. Although the PCFIA has a high-dose "hook" effect, this is of no analytical importance at concentrations of Mb less than or equal to 148,000 micrograms/L. The assay is not subject to interference from icterus (bilirubin less than or equal to 360 mg/L), has no cross-reaction with hemoglobin (less than or equal to 42 g/L), and may be performed with either plasma or serum in approximately 1 h. The intra- and interassay imprecisions (CV) of the method are less than 10% for concentrations of Mb within the normal range and less than 4% at higher concentrations. A comparison of the PCFIA with a commercial radioimmunoassay showed that results of the two assays correlate well (PCFIA = 0.88 x RIA + 18, r = 0.990, n = 171).

Detection of Antibody to Bordetella avium Using a Particle Concentration Fluorescence Immunoassay (PCFIA)

1991 ◽  
Vol 35 (4) ◽  
pp. 756 ◽  
Author(s):  
P. J. Blore ◽  
M. F. Slavik ◽  
N. K. Neighbor
Keyword(s):  
Particle Concentration ◽  
Fluorescence Immunoassay ◽  
Bordetella Avium

Particle concentration fluorescence immunoassay for measuring interleukin-6 receptor numbers

Cytokine ◽  
1991 ◽  
Vol 3 (1) ◽  
pp. 17-20 ◽  
Author(s):  
Zao-Dung Ling ◽  
Stephen Gillis ◽  
Liza J. Hart ◽  
David S. Matheson
Keyword(s):  
Interleukin 6 ◽  
Particle Concentration ◽  
Fluorescence Immunoassay ◽  
Interleukin 6 Receptor

Competitive particle concentration fluorescence immunoassay for measuring 5,10-dideaza-5,6,7,8-tetrahydrofolic acid (lometrexol) in serum

1991 ◽  
Vol 37 (2) ◽  
pp. 254-260 ◽  
Author(s):  
Larry D Taber ◽  
Peter O'Brien ◽  
Ronald R Bowsher ◽  
J Richard Sportsman
Keyword(s):  
Particle Concentration ◽  
Cross Reactivity ◽  
Minimum Detectable Concentration ◽  
Fluorescence Immunoassay ◽  
Standard Curve ◽  
Coefficients Of Variation ◽  
Analytical Recovery ◽  
Detectable Concentration ◽  
Tetrahydrofolic Acid ◽  
Cancer Agent

Abstract A competitive particle concentration fluorescence immunoassay (PCFIA) is described for measuring 5,10-dideaza-5,6,7,8-tetrahydrofolic acid (lometrexol; Lilly) in human serum. b-Phycoerythrin-labeled lometrexol competes with free lometrexol for binding to a limiting concentration of lometrexol-specific antibodies immobilized by a second antibody to submicrometer-diameter polystyrene particles in specially designed 96-well plates. Reaction particles are washed and concentrated onto filter membranes in the wells of the plates and the fluorescence is measured at 575 nm. The method, including sample preparation and data reduction, is automated and can be completed in less than 2 h. The assay has a standard curve maximum measurable concentration of 1000 micrograms/L and a minimum detectable concentration of 0.1 microgram/L. Analytical recovery of lometrexol in serum is quantitative at concentrations greater than 1 micrograms/L. Intra- and interassay coefficients of variation at 50 micrograms/L in serum are 7.1% (n = 9) and 7.5% (n = 33), respectively. The cross-reactivity of naturally occurring folates, folic acid analogs, and the anti-cancer agent methotrexate is minimal. We report the use of the PCFIA during Phase I clinical studies designed to evaluate the pharmacokinetics of lomextrexol after intravenous administration to cancer patients.

Particle concentration fluorescence immunoassay (PCFIA): a new, rapid immunoassay technique with high sensitivity

1984 ◽  
Vol 67 (1) ◽  
pp. 21-35 ◽  
Author(s):  
Michael E. Jolley ◽  
Chao-Huei J. Wang ◽  
Steven J. Ekenberg ◽  
Mark S. Zuelke ◽  
David M. Kelso
Keyword(s):  
Particle Concentration ◽  
High Sensitivity ◽  
Fluorescence Immunoassay ◽  
Immunoassay Technique

Rapid Direct Determination of Low and High Molecular Weight Kininogen in Human Plasma by Particle Concentration Fluorescence Immunoassay (PCFIA)

1997 ◽  
Vol 77 (01) ◽  
pp. 109-118 ◽  
Author(s):  
Cheryl F Scott ◽  
Bruce Shull ◽  
Werner Müller-Esterl ◽  
Robert W Colman
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Particle Concentration ◽  
Biological Fluids ◽  
Direct Determination ◽  
Plasma Samples ◽  
Fluorescence Immunoassay ◽  
Mean Values ◽  
Heavy Chains ◽  
Normal Humans

SummaryIn this study, we employed Particle Concentration Fluorescence Immunoassay (PCFIA), for directly measuring both high and low molecular weight kininogens (HK and LK) in human plasma. In 38 normal donors, the mean values for plasma kininogens were 93 mg/ml +/- 19 SD, 82 mg/ml +/- 12 SD, and 175 mg/ml +/- 29 SD, respectively for LK, HK, and TotK (the sum of LK and HK detected by their common heavy chains). Plasma completely deficient in HK and LK was unreactive (<0.25 mg/ml) in all 3 assays whereas plasma from a patient with Fitzgerald Trait had an HK value of 11 mg/ml, an LK value of 36 mg/ml and a TotK value of 59 mg/ml. The reagents can be prepared in advance and all three kininogen determinations can be performed, using the same diluted sample, on 24 plasma samples, in triplicate, or 40 plasma samples, in duplicate, in less than 1 h. By performing all 3 kininogen determinations, it is possible to differentiate cleaved from intact kininogens. This technique will facilitate the widespread screening of kininogen levels in biological fluids of normal humans as well as of patients with various diseases.

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