Light-chain ratios of immunoglobulins G, A, and M determined by enzyme immunoassay

1990 ◽  
Vol 36 (3) ◽  
pp. 501-502 ◽  
Author(s):  
S H Chui ◽  
C W Lam ◽  
K N Lai
Keyword(s):  
Hydrogen Peroxide ◽  
Detection Limit ◽  
Light Chain ◽  
Reference Intervals ◽  
Light Chains ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microtiter Plates ◽  
Urea Hydrogen Peroxide ◽  
Apparently Healthy

Abstract We describe an enzyme-linked immunosorbent assay for determination of light-chain ratios for IgG, IgA, and IgM in serum. A commercial serum with known overall kappa and lambda concentrations was used as standard. To capture the respective immunoglobulins, we used antibodies to gamma, lambda and mu chains coated onto microtiter plates. Peroxidase-conjugated anti-kappa and anti-lambda chain antisera were reacted with light chains on the captured immunoglobulins, and the amount of enzyme bound was monitored with o-phenylenediamine and urea-hydrogen peroxide as substrates. Calculation of absorbance ratios allowed determination of kappa and lambda chain concentrations of individual immunoglobulins in the standard and samples. Within-run and between-run CVs (n = 25) ranged from 5.9% to 13.0% for "high," "normal," and "low" kappa/lambda ratios for IgG, IgA, and IgM. The thoroughness of light-chain detection, expressed as, e.g., (IgA kappa + IgA lambda)/(total IgA), for 150 sera was 91-110%. The detection limit was 1 microgram/L. Reference intervals (mean +/- SD) for kappa/lambda ratios in sera from 100 apparently healthy adults were 2.34 +/- 0.80 for IgG, 1.59 +/- 0.40 for IgA, and 1.86 +/- 0.76 for IgM.

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.

Rapid determination of the hypoxanthine increase in ischemic exercise tests.

1988 ◽  
Vol 34 (8) ◽  
pp. 1607-1610 ◽  
Author(s):  
P A Bolhuis ◽  
R Zwart ◽  
P R Bär ◽  
M de Visser ◽  
H J van der Helm
Keyword(s):  
Hydrogen Peroxide ◽  
Detection Limit ◽  
Rapid Determination ◽  
Serum Lactate ◽  
Benzenesulfonic Acid ◽  
Exercise Tests ◽  
Ph Optimum ◽  
Absorbance Maximum ◽  
Myoadenylate Deaminase

Abstract After ischemic exercise tests, performed to detect glycogenoses or myoadenylate deaminase (EC 3.5.4.6) deficiency, the increases in serum lactate and ammonia usually are measured. Determination of hypoxanthine instead of ammonia can also be used to show myoadenylate deaminase deficiency, but HPLC of hypoxanthine is time-consuming. As a substitute, we developed an indirect enzymatic equilibrium method for hypoxanthine based on coupling the chromogenic system 3,5-dichloro-2-hydroxy-benzenesulfonic acid/4-aminophenazone with formation of hydrogen peroxide by xanthine oxidase (EC 1.1.3.22). The pH optimum is at 7.8 and the absorbance maximum at 510 nm. The calibration curve is linear from 0 to 100 mumol/L and the detection limit is 0.9 mumol/L. Analytical variability (CV) was 1.5% to 3.6% within-run, 4.5% to 8.5% between-run. The assay can be performed with a standard spectrophotometer or a centrifugal analyzer. The coefficient of correlation was 0.68 between hypoxanthine and ammonia increases in plasma from controls who performed the exercise test.

scholarly journals Serum Reference Intervals and Diagnostic Ranges for Free κ and Free λ Immunoglobulin Light Chains: Relative Sensitivity for Detection of Monoclonal Light Chains

2002 ◽  
Vol 48 (9) ◽  
pp. 1437-1444 ◽  
Author(s):  
Jerry A Katzmann ◽  
Raynell J Clark ◽  
Roshini S Abraham ◽  
Sandra Bryant ◽  
James F Lymp ◽  
...  
Keyword(s):  
Light Chain ◽  
Reference Intervals ◽  
Systemic Amyloidosis ◽  
Light Chains ◽  
Light Chain Deposition Disease ◽  
Monoclonal Gammopathies ◽  
Automated Immunoassay ◽  
Light Chain Deposition ◽  
Diagnostic Intervals ◽  
Detection And Quantification

Abstract Background: The detection of monoclonal free light chains (FLCs) is an important diagnostic aid for a variety of monoclonal gammopathies and is especially important in light-chain diseases, such as light-chain myeloma, primary systemic amyloidosis, and light-chain-deposition disease. These diseases are more prevalent in the elderly, and assays to detect and quantify abnormal amounts of FLCs require reference intervals that include elderly donors. Methods: We used an automated immunoassay for FLCs and sera from a population 21–90 years of age. We used the calculated reference and diagnostic intervals to compare FLC results with those obtained by immunofixation (IFE) to detect low concentrations of monoclonal κ and λ FLCs in the sera of patients with monoclonal gammopathies. Results: Serum κ and λ FLCs increased with population age, with an apparent change for those >80 years. This trend was lost when the FLC concentration was normalized to cystatin C concentration. The ratio of κ FLC to λ FLC (FLC K/L) did not exhibit an age-dependent trend. The diagnostic interval for FLC K/L was 0.26–1.65. The 95% reference interval for κ FLC was 3.3–19.4 mg/L, and that for λ FLC was 5.7–26.3 mg/L. Detection and quantification of monoclonal FLCs by nephelometry were more sensitive than IFE in serum samples from patients with primary systemic amyloidosis and light-chain-deposition disease. Conclusions: Reference and diagnostic intervals for serum FLCs have been developed for use with a new, automated immunoassay that makes the detection and quantification of monoclonal FLCs easier and more sensitive than with current methods. The serum FLC assay complements IFE and allows quantification of FLCs in light-chain-disease patients who have no detectable serum or urine M-spike.

Monoclonal antibody-based solid-phase immunoenzymometric assays for quantifying human immunoglobulin G and its subclasses in serum.

1985 ◽  
Vol 31 (12) ◽  
pp. 1940-1945 ◽  
Author(s):  
C Papadea ◽  
I J Check ◽  
C B Reimer
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Solid Phase ◽  
Detection System ◽  
Reference Intervals ◽  
Microtiter Plates ◽  
Myeloma Proteins ◽  
Human Proteins ◽  
Reference Preparation ◽  
Apparently Healthy

Abstract We developed quantitative immunoenzymometric assays for human IgG and its subclasses by using monoclonal antibodies, an avidin-biotin detection system and, as the calibrant, the U.S. National Reference Preparation for Specific Human Proteins. The assays are sensitive (detecting as little as 6 micrograms/L), precise (average inter-assay CV less than 11%), and vary linearly with concentrations over a five- to 10-fold range, depending on the monoclonal antibody. We evaluated 22 different monoclonal antibodies, many of which remained highly reactive when immobilized in wells of microtiter plates coated with bovine serum albumin-glutaraldehyde to "capture" total IgG or subclasses of IgG in the sample. We demonstrated the specificity of the most reactive antibodies by using a panel of 20 purified myeloma proteins. The sum of IgG subclass concentrations correlated well (r = 0.84, p less than 0.001) with the total IgG measured in sera from 63 apparently healthy adults (26 men, 37 women). We estimated 95 percentile reference intervals for the immunoglobulins in these subjects and determined the following mean percentage distributions of IgG subclasses: IgG1 49, IgG2 33, IgG3 9, and IgG4 7. The availability of these assays should facilitate studies of the clinical significance of the subclasses.

Development of an Analytical Protocol for the Determination of Polybrominated Biphenyls in Water by Enzyme-Linked Immunosorbent Assay

2011 ◽  
Vol 347-353 ◽  
pp. 1537-1541 ◽  
Author(s):  
Han Yu Chen ◽  
Hui Sheng Zhuang
Keyword(s):  
Detection Limit ◽  
Quantitative Determination ◽  
Incubation Time ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Concentration ◽  
Polybrominated Biphenyls ◽  
Analytical Protocol ◽  
Optimized Conditions

A novel immunoassay has been developed for the quantitative determination of polybrominated biphenyls (PBBs) using indirect competitive format. A new method was developed to synthesize PBBs congener (PCB15) hapten and its artificial immunogen, then the polyclonal antibodies. The assay was optimized concerning the coating conjugate and antibody concentration, incubation time and temperature, the tolerance to organic solvents and so on. Under optimized conditions, PBB15 can be determined in the concentration range of 0.01-100 μg L-1 with a detection limit of 0.02 μg L-1. The cross-reactivities of the assays were below 8%. While water samples could be analyzed directly.

Determination of dehydroepiandrosterone sulfate in plasma by a one-step enzyme immunoassay with a microtitre plate.

1985 ◽  
Vol 31 (11) ◽  
pp. 1876-1879 ◽  
Author(s):  
T K Dhar ◽  
C Müller ◽  
M Schöneshöfer
Keyword(s):  
Hydrogen Peroxide ◽  
Detection Limit ◽  
Enzyme Immunoassay ◽  
Cost Effective ◽  
Dehydroepiandrosterone Sulfate ◽  
Turnaround Time ◽  
Microtitre Plate ◽  
Standard Curve ◽  
One Step

Abstract We have developed a rapid and cost-effective enzyme immunoassay for dehydroepiandrosterone sulfate (DHEA-S) in plasma, performed with samples on a microtitre plate within 2.5 h. No extraction or centrifugation steps are involved. The 3-hemisuccinate of dehydroepiandrosterone is labeled with horseradish peroxidase, then mixed with hydrogen peroxide substrate in the presence of the chromogen, tetramethylbenzidine. The detection limit of the assay is 12.5 pg of DHEA-S per well. Intra- and interassay CVs at three steroid concentrations (12.8, 1.28, and 0.16 mumol/L) ranged from 2.3 to 5.4% and 6.1 to 8.4%, respectively. Results correlated well (r = 0.95) with those of a radioimmunoassay with iodinated DHEA-S. The turnaround time for 41 samples (in duplicate) is 2.5 h, which includes 2 h of incubation time. The sensitivity of this one-step version and the linearity of its standard curve are equivalent to those of a less practicable two-step version. This technique may replace coated-tube enzyme immunoassays for routine use.

scholarly journals An Immunoaffinity Column for the Determination of Peanut Protein in Chocolate

1999 ◽  
Vol 82 (3) ◽  
pp. 666-668 ◽  
Author(s):  
Harvey W Newsome ◽  
Michael Abbott
Keyword(s):  
Detection Limit ◽  
Immunoaffinity Chromatography ◽  
Polyclonal Antiserum ◽  
Peanut Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunoaffinity Column ◽  
Rabbit Polyclonal Antiserum ◽  
The Matrix ◽  
Direct Elisa

Abstract An immunoaffinity column was prepared from rabbit polyclonal antiserum for the determination of peanut protein from food matrixes. The anti-peanut immunoglobulin G was isolated from antiserum by affinity chromatography on a column coupled with peanut protein and then attached to an AminoLink gel. The column was applied to the determination of peanut protein in chocolate after extraction, immunoaffinity chromatography, and enzyme-linked immunosorbent assay (ELISA). Overall recoveries from chocolate spiked with 0.2–3.2 μg/g of peanut protein averaged 77% (range, 72–84%), and the minimum detection limit was 0.1 μg/g. Chromatography of extracts with the column improved detection limit and eliminated the matrix effect experienced with direct ELISA of chocolate extracts.

scholarly journals Determination of Egg Proteins in Food Products by Enzyme Immunoassay

2000 ◽  
Vol 83 (1) ◽  
pp. 139-143 ◽  
Author(s):  
Jupiter M Yeung ◽  
W Harvey Newsome ◽  
Michael A Abbott
Keyword(s):  
Detection Limit ◽  
Enzyme Immunoassay ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antigen Binding ◽  
Food Ingredients ◽  
Coefficients Of Variation ◽  
Egg Products ◽  
Whole Egg

Abstract An enzyme-linked immunosorbent assay (ELISA) was developed to determine the presence of egg proteins in foods. The polyclonal antibodies developed were specific to whole egg proteins and did not cross-react with any of the 38 nuts, legumes, or other common food ingredients tested. The concentrations of egg proteins that will inhibit 50% of antibody–antigen binding, IC50, were 3–7 ng/mL, and the linear range was 0.5–62.5 ng/mL. The detection limit was 0.2 ppm for various foods. Recoveries ranged from 67 to 96%. The intra- and inter-assay coefficients of variation in this procedure were 10–13% for ice cream spiked at 0.8 and 1.6 ppm. The ELISA has been applied to ice creams, noodles, pasta, and breads. Egg proteins were identified in all declared egg products, and no false positives were found.

Pediatric and Perinatal Reference Intervals for Immunoglobulin Light Chains Kappa and Lambda

1992 ◽  
Vol 38 (4) ◽  
pp. 548-550 ◽  
Author(s):  
K R Herkner ◽  
H Salzer ◽  
A Böck ◽  
A Mühl ◽  
T Tsaka ◽  
...  
Keyword(s):  
Light Chain ◽  
Reference Intervals ◽  
Linear Increase ◽  
Light Chains ◽  
Serum Concentrations ◽  
Immunoglobulin Light Chains ◽  
Adult Type ◽  
Mean Values ◽  
Age Related ◽  
Chain Analysis

Abstract In routine analysis for immunoglobulin light chains in pediatric diagnostics, the age-related reference intervals for serum kappa (kappa) and lambda (lambda) light chains were evaluated in 1543 healthy subjects (newborns to age 16 years, including 168 premature infants). Light-chain analysis was performed by rate nephelometry. IgG, IgA, and IgM were measured simultaneously, and heavy- and light-chain differences were calculated for control purposes. Results for IgG, IgA, and IgM generally agreed with reference intervals reported in the literature. kappa showed age-related changes comparable with changes in IgG concentrations, whereas lambda showed moderate fluctuations. The kappa/lambda ratio showed an almost linear increase with age, starting with 0.97 at four months and reaching the highest value of 2.21 at 15 years (mean values). Preterm infants presented with markedly low serum concentrations of IgG and corresponding light chains but with adult-type kappa/lambda ratios because of the maternal-origin IgG.

Oxidation Reactions Catalyzed by Polyoxomolybdate Salts

2013 ◽  
Vol 68 (5-6) ◽  
pp. 587-597 ◽  
Author(s):  
Bo Zhang ◽  
Su Li ◽  
Alexander Pöthig ◽  
Mirza Cokoja ◽  
Shu-Liang Zang ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Environmentally Benign ◽  
Oxidation Reactions ◽  
X Ray Diffraction ◽  
Oxidation Of Sulfides ◽  
Urea Hydrogen Peroxide ◽  
Ionic Compounds ◽  
Solid State Structures ◽  
Epoxidation Of Olefins

Ionic compounds containing the polyoxomolybdate anion [Mo6O19]2- and [(n-C4H9)4P]+ (tetrabutylphosphonium), [(n-C4H9)3P(n-C14H29)]+ (tributyl (tetradecyl)phosphonium), [Bmim]+ (1- butyl-3-methylimidazolium) and [Dbmim]+ (1,2-dimethyl-3-butylimidazolium) cations were prepared and characterized, including the determination of three of the solid state structures by singlecrystal X-ray diffraction. These compounds were applied as catalysts for the epoxidation of olefins with urea hydrogen peroxide (UHP) as oxidant in the ionic liquid [Bmim]PF6. Additionally, the oxidation of sulfides to sulfoxides with hydrogen peroxide (H2O2) in several solvents was investigated. The polyoxomolybdate catalysts showed a good performance for epoxidation of olefins as well as for oxidation of sulfides. Furthermore, the catalysts can be recycled several times in oxidation reactions. We present this methodology for the oxidation reaction in a simple, economically, technically, and environmentally benign manner

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