scholarly journals Direct Assay for Cobalamin Bound to Transcobalamin (Holo-Transcobalamin) in Serum

2002 ◽  
Vol 48 (3) ◽  
pp. 526-532 ◽  
Author(s):  
Marius Ulleland ◽  
Ingar Eilertsen ◽  
Edward V Quadros ◽  
Sheldon P Rothenberg ◽  
Sergey N Fedosov ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Quantitative Determination ◽  
Reliable Method ◽  
Solid Phase ◽  
Reference Interval ◽  
Magnetic Microspheres ◽  
Affinity Constant ◽  
Capture Assay

Abstract Background: Only cobalamin carried by transcobalamin (holo-transcobalamin) is available for cellular uptake and hence is physiologically relevant. However, no reliable or accurate methods for quantifying holo-transcobalamin are available. We report a novel holo-transcobalamin assay based on solid-phase capture of transcobalamin. Methods: A monoclonal antibody specific for human transcobalamin with an affinity constant >1010 L/mol was immobilized on magnetic microspheres to capture and concentrate transcobalamin. The cobalamin bound to transcobalamin was then released and assayed by a competitive binding radioassay. The quantification of holo-transcobalamin was accomplished using calibrators composed of recombinant, human holo-transcobalamin. Results: The assay was specific for holo-transcobalamin and had a detection limit of 5 pmol/L. Within-run and total imprecision (CV) was 5% and 8–9%, respectively. The working range (CV <20%) was 5–370 pmol/L. Dilutions of serum were linear in the assay range. The recovery of recombinant, human holo-transcobalamin added to serum was 93–108%. A 95% reference interval of 24–157 pmol/L was established for holo-transcobalamin in 105 healthy volunteers 20–80 years of age. For 72 of these sera, holo-haptocorrin and total cobalamin were also determined. Whereas holo-haptocorrin correlated well (r2 = 0.87) with total cobalamin, holo-transcobalamin correlated poorly (r2 = 0.23) with total cobalamin or holo-haptocorrin. Conclusions: The solid-phase capture assay provides a simple, reliable method for quantitative determination of holo-transcobalamin in serum.

scholarly journals Determination of Haptoglobin in Bovine Serum using Polyclonal and Monoclonal Anti-human Haptoglobin Antibodies

2010 ◽  
Vol 79 (1) ◽  
pp. 105-112 ◽  
Author(s):  
Paulina Jawor ◽  
Tadeusz Stefaniak ◽  
Iwona Kątnik-Prastowska
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Bovine Serum ◽  
Solid Phase ◽  
Low Cost ◽  
Reference Method ◽  
Serum Samples ◽  
Incubation Periods ◽  
Short Time

Two ELISA procedures to determine haptoglobin (Hp) in bovine serum were developed. Equine haemoglobin was used as the solid phase. Self-developed goat polyclonal antibody (variant I) and monoclonal antibody (variant II) raised against human Hp were used. The results were compared with the guaiacol method. High correlation was found (r = 0.96 and r = 0.90, respectively) based on the results of 548 bovine serum samples, of which 357 were from clinically healthy cows and 191 from cows and calves monitored during treatment for the most common diseases. The Hp detection limit of ELISA using polyclonal Ab was 0.1 mg/l and using MoAb 0.21 mg/l. The addition of 2% PEG 6000 at the antibody-binding steps enabled major shortening of the incubation periods. The relatively short time, low cost of reagents, and high correlation with the reference method support the use of these ELISA variants in bovine diagnostics.

scholarly journals Quantitative Determination of Acetamiprid in Pollen Based on a Sensitive Enzyme-Linked Immunosorbent Assay

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1265 ◽  
Author(s):  
Qingkui Fang ◽  
Quan Zu ◽  
Xiude Hua ◽  
Pei Lv ◽  
Wanwen Lin ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Quantitative Determination ◽  
Immunosorbent Assay ◽  
Linear Range ◽  
Enzyme Linked Immunosorbent Assay ◽  
Experimental Conditions ◽  
Coating Antigen ◽  
Relative Standard

A sensitive biotinylated indirect competitive enzyme-linked immunosorbent assay (Bic-ELISA) was developed to detect acetamiprid pesticides in pollen, based on the heterogeneous coating antigen and biotinylated anti-acetamiprid monoclonal antibody. Under optimized experimental conditions, the detection limit for the Bic-ELISA was 0.17 ng/mL and the linear range was 0.25–25 ng/mL. The cross-reactivities could be regarded as negligible for the biotinylated antibodies with their analogues except for thiacloprid (1.66%). Analyte recoveries for extracts of spiked pollen (camellia pollen, lotus pollen, rape pollen) ranged from 81.1% to 108.0%, with intra-day relative standard deviations (RSDs) of 4.8% to 10.9%, and the average reproducibility was 85.4% to 110.9% with inter-assay and inter-assay RSDs of 6.1% to 11.7%. The results of Bic-ELISA methods for the Taobao’s website samples were largely consistent with HPLC-MS/MS. Therefore, the established Bic-ELISA methods would be conducive to the monitoring of acetamiprid in pollen.

Liquid Chromatographic Determination of Sulfamoyldapsone in Swine Tissues

1987 ◽  
Vol 70 (6) ◽  
pp. 1031-1032
Author(s):  
Yuuko S Endoh ◽  
Ryozo Yamaoka ◽  
Nobuo Sasaki
Keyword(s):  
Detection Limit ◽  
Quantitative Determination ◽  
Chromatographic Determination ◽  
Alumina Column ◽  
Average Recovery ◽  
Diaminodiphenyl Sulfone ◽  
Alumina Column Chromatography ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract A liquid chromatographic (LC) method is described for the quantitative determination of sulfamoyldapsone (2-sulfamoyl-4,4'-diaminodiphenyl sulfone) in swine muscle, liver, kidney, and fat. Sulfamoyldapsone was extracted from tissues with acetonitrile saturated with n-hexane. The extract was washed with n-hexane saturated with acetonitrile, concentrated, and cleaned up by alumina column chromatography. Sulfamoyldapsone was separated on an ODS column by using acetonitrile-methanol-water (6 + 18 + 76) and was detected at 292 nm. Overall average recovery of sulfamoyldapsone added to tissues at levels of 0.1 and 0.5 /μg/g was 93.3% ± 6.0. Detection limit was 0.02 μg/g in these tissues.

Direct solid-phase enzymoimmunoassay of testosterone in saliva.

1989 ◽  
Vol 35 (10) ◽  
pp. 2044-2047 ◽  
Author(s):  
K Howard ◽  
M Kane ◽  
A Madden ◽  
J P Gosling ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Medical Applications ◽  
Microtiter Plates ◽  
Large Numbers ◽  
Coefficients Of Variation ◽  
Serial Sampling ◽  
Direct Solid

Abstract This competitive, solid-phase enzymoimmunoassay for testosterone in saliva is carried out on microtiter plates and involves no chromatographic or extraction steps. With an overnight incubation the detection limit of the assay is 230 fg per well (16.1 pmol/L). There was a good correlation (correlation coefficient 0.95) between testosterone concentrations measured with and without prior extraction of the saliva samples. Repeated assay of three control saliva samples containing a range of testosterone concentrations (200-1000 pmol/L) gave within- and between-assay coefficients of variation of 5.5-13.2%. The analytical procedure is simple and closely resembles already published procedures for the determination of progesterone and estrone (with extraction) in saliva. One person can assay 200 samples in 24 h and the assay is suitable for reproductive and sports medical applications, particularly for projects involving serial sampling and yielding large numbers of samples.

scholarly journals Preconcentration of Copper with Solid Phase Extraction and its Determination by Flame Atomic Absorption Spectrometry

2008 ◽  
Vol 5 (4) ◽  
pp. 878-883 ◽  
Author(s):  
Ayob Parchehbaf Jadid ◽  
Habibollah Eskandari
Keyword(s):  
Detection Limit ◽  
Solid Phase Extraction ◽  
Solid Phase ◽  
Copper Ions ◽  
Absorption Spectrometry ◽  
Phase Extraction ◽  
Flame Atomic Absorption ◽  
Low Levels ◽  
Determination Of Copper

A new method for the solid phase extraction (SPE) and determination of copper ions at low levels is presented. Extraction percent and the effects of some factors were evaluated. The detection limit was in the range of 2.26 µg·L-1. This procedure has been successfully applied to determination of copper in water samples.

Solid-phase enzymoimmunoassay for osteocalcin in human serum or plasma, with use of a monoclonal antibody.

1989 ◽  
Vol 35 (10) ◽  
pp. 2087-2092 ◽  
Author(s):  
M J Power ◽  
P F Fottrell
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Horseradish Peroxidase ◽  
Human Serum ◽  
Solid Phase ◽  
Sample Volume ◽  
Microtiter Plates ◽  
Analytical Recovery ◽  
Peroxidase Conjugate

Abstract In this solid-phase enzymoimmunoassay on microtiter plates for osteocalcin in serum or plasma, we use an osteocalcin-horseradish-peroxidase conjugate and a monoclonal antibody raised against bovine osteocalcin. We thoroughly standardized the assay for measurement of osteocalcin in both serum and plasma, demonstrating independence of sample volume, and determining the analytical recovery and within-and between-assay CVs. The detection limit was between 0.6 and 1.1 micrograms/L and the ED50 was 16 micrograms/L for a 5-microL sample volume. The intra-assay CV over the range 3 to 74 micrograms/L was less than or equal to 15%. The interassay CV over the range 3.6 to 46 micrograms/L was less than or equal to 16%. Results by this assay and by an in-house radioimmunoassay in which the same monoclonal antibody was used correlated well (r2 = 0.948). Osteocalcin concentrations in serum and plasma as measured with the present assay agreed well with published values.

scholarly journals Solid-Phase Extraction and Gas Chromatography with Mass Spectrometric Determination of Capsaicin and Some of Its Analogues from Chili Peppers (Capsicum spp.)

1999 ◽  
Vol 82 (6) ◽  
pp. 1399-1405 ◽  
Author(s):  
Philemon Manirakiza ◽  
Adrian Covaci ◽  
Paul Schepens
Keyword(s):  
Gas Chromatography ◽  
Quantitative Determination ◽  
Solid Phase ◽  
Accurate Method ◽  
Mass Spectrometric ◽  
Phase Extraction ◽  
Capsicum Spp ◽  
The Mean ◽  
Chili Peppers

Abstract A rapid and accurate method has been developed for the quantitative determination of capsaicin and its most important analogues, dihydrocapsaicin and nordihydrocapsaicin in chili peppers. These components were extracted with methylene chlo ride and separated from interfering substances with activated charcoal. Further cleanup on Florisil cartridges and elution with ethyl acetate were performed before gas chromatographic with mass spectrometric quantitation. The concentrations found were 440 ± 64 μg/g capsaicin, 81 ± 10 μg/g dihydrocapsaicin, and 11 ± 2 μg/g nordihydrocapsaicin. The mean recovery values for triplicate analysis were between 85-94%.

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