Nonspecific binding as a source of error in thyrotropin radioimmunoassay with polyethylene glycol as separating agent.

1980 ◽  
Vol 26 (3) ◽  
pp. 487-490
Author(s):  
I W Chen ◽  
L Heminger ◽  
H R Maxon ◽  
J Y Tsay
Keyword(s):  
Polyethylene Glycol ◽  
Specific Binding ◽  
Mean Value ◽  
Total Serum Protein ◽  
Total Serum ◽  
Albumin Concentration ◽  
Nonspecific Binding ◽  
Antibody Complex ◽  
The Individual ◽  
Antigen Antibody

Abstract We investigated the effects of nonspecific binding on thyrotropin values obtained by radioimmunoassay in which polyethylene glycol is used as precipitant. Differences in nonspecific binding among individual samples were significant (F-test, p less than 0.001, range 5.5 to 14.1%). Non-specific binding and total serum protein were directly correlated (r = 0.472, n = 59; p less than 0.001). Nonspecific binding increased with increasing concentrations of globulins but showed no relation to albumin concentration. If globulin concentration was less than 15 g/L, precipitation of the antigen—antibody complex by polyethylene glycol was incomplete. The mean value for thyrotropin in sera from 67 healthy subjects was 2.7 (SD 0.3) milli-international units per liter (milli-int. unit/L) without individual serum nonspecific binding correction, significantly (p less than 0.005) higher than that with nonspecific binding correction (1.6, SD 0.1, milli-int. unit/L). Evidently, inter-sample variations in nonspecific binding may cause significant errors under these conditions, which can be minimized by taking into account the individual nonspecific binding of each serum sample.

Nonspecific binding as a source of error in thyrotropin radioimmunoassay with polyethylene glycol as separating agent.

1980 ◽  
Vol 26 (3) ◽  
pp. 487-490 ◽  
Author(s):  
I W Chen ◽  
L Heminger ◽  
H R Maxon ◽  
J Y Tsay
Keyword(s):  
Polyethylene Glycol ◽  
Specific Binding ◽  
Mean Value ◽  
Total Serum Protein ◽  
Total Serum ◽  
Albumin Concentration ◽  
Nonspecific Binding ◽  
Antibody Complex ◽  
The Individual ◽  
Antigen Antibody

Abstract We investigated the effects of nonspecific binding on thyrotropin values obtained by radioimmunoassay in which polyethylene glycol is used as precipitant. Differences in nonspecific binding among individual samples were significant (F-test, p less than 0.001, range 5.5 to 14.1%). Non-specific binding and total serum protein were directly correlated (r = 0.472, n = 59; p less than 0.001). Nonspecific binding increased with increasing concentrations of globulins but showed no relation to albumin concentration. If globulin concentration was less than 15 g/L, precipitation of the antigen—antibody complex by polyethylene glycol was incomplete. The mean value for thyrotropin in sera from 67 healthy subjects was 2.7 (SD 0.3) milli-international units per liter (milli-int. unit/L) without individual serum nonspecific binding correction, significantly (p less than 0.005) higher than that with nonspecific binding correction (1.6, SD 0.1, milli-int. unit/L). Evidently, inter-sample variations in nonspecific binding may cause significant errors under these conditions, which can be minimized by taking into account the individual nonspecific binding of each serum sample.

scholarly journals Blood Biochemical of Nile Crocodile (Crocodylus niloticus) in Kano Zoological Garden, Nigeria

2019 ◽  
Vol 2 (1) ◽  
pp. 31-37
Author(s):  
Adelakun K.M. ◽  
Kehinde A.S. ◽  
Laoye O. ◽  
Ihidero A.A. ◽  
Dalha A.
Keyword(s):  
Density Lipoprotein ◽  
Clinical Pathology ◽  
Clinical Information ◽  
Cholesterol Concentration ◽  
Total Serum Protein ◽  
Total Serum ◽  
Breakdown Product ◽  
Albumin Concentration ◽  
Zoological Garden ◽  
Nile Crocodile

The potential application of blood reference range for crocodile is a basis that can provide important clinical information about health and physiological condition of the animal. This study investigates serum biochemistry of Nile crocodile from Kano Zoological Garden, Kano, Nigeria. Six (6) adult Nile crocodile (Crocodylus niloticus) were captured from crocodile pond in the zoo. Blood was collected from post-occipital sinus of the physically restrain crocodile and used for serum biochemical parameters. The results revealed the Total Serum Protein (TSP) concentration of 9.2g/Ɩ, albumin concentration which is a common plasma protein is 43g/Ɩ while globulin concentration is 54g/Ɩ. Cholesterol concentration measure is registered at 5.2mmol/Ɩ with High-Density Lipoprotein (HDL) and Low-Density Lipoprotein (LDL) of 1mmol/Ɩ and 1.35mmol/Ɩ respectively. Creatinine: a breakdown product of creatinine which is an important part of muscle tissue is 44umol/Ɩ. Uric acid which is a primary catabolic end product of protein is 0.18mmol/Ɩ while glucose and triglyceride are 4.94mmol/Ɩ and 2.24mmol/Ɩ respectively while enzymes which include Alanine aminotransferase (ALT) concentration is 6U/Ɩ, Aspartate aminotransferase concentration is 5U/Ɩ while Alkaline Phosphatase is 20U/Ɩ. The biochemical values recorded were compared with available data on farm Nile crocodile. Clearly, nutritional status, age, gender, season, physiology and environment should be considered if clinical pathology is to be employed as a diagnostic tool.

Serum glycoproteins in growing pups

1962 ◽  
Vol 203 (1) ◽  
pp. 119-121 ◽  
Author(s):  
Otakar V. Sirek ◽  
Anna Sirek
Keyword(s):  
Weight Gain ◽  
Sialic Acid ◽  
Serum Protein ◽  
Total Protein ◽  
Protein Concentration ◽  
Total Serum Protein ◽  
Total Serum ◽  
Albumin Fraction ◽  
The Individual ◽  
Serum Protein Concentration

Total protein-bound hexose, hexosamine, and sialic acid were determined in sera of six littermate mongrel pups at monthly intervals from the 4th day after birth up to the age of 7 months. The concentration of the individual constituents fluctuated considerably from month to month, but the values showed neither a definite trend nor a relationship to weight gain. When the carbohydrate moiety was expressed as percentage of total serum protein concentration, the values were high in newborn pups and diminished after the 1st month of life. This was due to a rise in the concentration of total serum protein, brought about by an increase of the albumin fraction which is low in carbohydrate.

An improved radioimmunoassay of C-peptide and its application in a multiyear study.

1989 ◽  
Vol 35 (1) ◽  
pp. 37-42 ◽  
Author(s):  
J E Myrick ◽  
E W Gunter ◽  
V L Maggio ◽  
D T Miller ◽  
W H Hannon
Keyword(s):  
Nonspecific Binding ◽  
C Peptide ◽  
Freeze Thaw ◽  
Serum Pool ◽  
Hands On ◽  
Antibody Complex ◽  
Analytical Range ◽  
Antigen Antibody ◽  
Free Serum ◽  
Differential Precipitation

Abstract A commercial radioimmunoassay (RIA) for human proinsulin C-peptide was modified to improve its ruggedness and specificity, to decrease the influence of specimen matrix, and to shorten "hands-on" time. In the new protocol, we prepare calibrators in a C-peptide-free serum pool, prepared by treatment with activated charcoal (biological matrix), instead of in a defined matrix. This yielded essentially 100% analytical recoveries for C-peptide concentrations up to 300 pmol/L, a broader analytical range. We also corrected calibrators and unknown samples for nonspecific binding (NSB). Decreasing the concentration of ethanol (from 950 to 880 mL/L) for differential precipitation of the antigen-antibody complex resulted in an NSB of less than 10%, while maintaining high bound/total count percentages for samples and calibrators. C-peptide is thermally unstable without aprotinin at -20 degrees C and with or without aprotinin at 4 degrees C or above, but multiple freeze-thaw cycles do not affect C-peptide in serum. The modified C-peptide assay was applied to plasma from a multiyear study (fasting and post-carbohydrate-challenge subjects). During the four years of the study CVs ranged from 1.9% to 8.6% for replicate analyses of C-peptide in samples with concentrations less than or equal to 500 pmol/L. Between-run CVs were 3.8% to 8.2%, total CVs 3.8% to 10.7%.

Real-Time Detection of Antigen–Antibody Reactions by Imaging Ellipsometry

2007 ◽  
Vol 60 (9) ◽  
pp. 667 ◽  
Author(s):  
Irina Chamritski ◽  
Mark Clarkson ◽  
Jeff Franklin ◽  
Shi Wei Li
Keyword(s):  
Real Time ◽  
Specific Binding ◽  
Protein A ◽  
Surface Proteins ◽  
Human Antibody ◽  
Label Free ◽  
Time Resolved ◽  
Imaging Ellipsometry ◽  
Antibody Complex ◽  
Antigen Antibody

In the field of proteomics the quantification of the affinity of an antibody to its partners and the evaluation of its specific binding is an important issue. With an imaging ellipsometer the interaction of an antibody with immobilized antigens on a model microarray is observed in a time-resolved and label-free manner. Imaging ellipsometry was developed for real-time monitoring of the biomolecule interaction between an antigen in solution and an antibody immobilized on a silicon surface. Proteins were immobilized by the formation of carboxy-alkyl monolayers on silicon substrates, where a biotin-labelled antibody was immobilized by a biotin–streptavidin linkage. Anti-human IgG bound specifically to human antibody and protein A, similarly anti-goat IgG bound to goat antibody. No binding was observed between anti-rabbit IgG and goat antibody. All stages of the formation of the antigen–antibody complex were imaged by imaging ellipsometry. By monitoring changes in y, the mole fraction θ of the antigen–antibody binding was determined. Immunological reactions of two different antigen–antibody combinations were fitted by the Langmuir adsorption equation, and affinity constants for two reactions were calculated.

scholarly journals The relationship between an elevated serum amino acid ratio and the development of other biological abnormalities in the experimentally malnourished pig

1969 ◽  
Vol 23 (4) ◽  
pp. 791-804 ◽  
Author(s):  
R. F. Grimble ◽  
R. G. Whitehead
Keyword(s):  
Amino Acid ◽  
Serum Protein ◽  
Elevated Serum ◽  
Total Serum Protein ◽  
Total Serum ◽  
Albumin Concentration ◽  
Acid Ratio ◽  
Total Calorie ◽  
The Relationship ◽  
Deficient Group

1. Weanling pigs were fed under three dietary regimens, control, low protein and total calorie restricted.2. In the protein-deficient group the amino acid ratio did not start to become elevated until growth was impaired and total serum protein and albumin concentration began to fall.3. In the protein-deficient group, but not in the control or undernourished animals, the magnitude of the ratio was statistically correlated with the rate of growth, appetite, serum protein and albumin concentration and hydroxyproline excretion.4. The results provide information on the relationship between the serum amino acid ratio and nutritional status.

Two-hour assay for lutropin during ovulation.

1981 ◽  
Vol 27 (5) ◽  
pp. 727-730 ◽  
Author(s):  
D L Hay ◽  
P A Tasker ◽  
W I Johnston ◽  
I Horacek
Keyword(s):  
Polyethylene Glycol ◽  
Immune Complex ◽  
Incubation Time ◽  
Equilibrium Concentration ◽  
Optimal Conditions ◽  
Nonspecific Binding ◽  
Low Concentrations ◽  
Assay Time ◽  
High Concentrations ◽  
Antigen Antibody

Abstract A rapid lutropin assay with a 2-h incubation time and a second antibody/polyethylene glycol separation step is presented. Assay time is shortened by incubating at 37 degrees C and using relatively high concentrations of antibody and radioligand. The interval required for the antigen/antibody reaction varies directly with lutropin concentration, from 1 h for ovulation values to 8 h for low values. After a 2-h incubation, low concentrations have reached 80% of their equilibrium concentration. Separation of the bound fraction by use of combined second antibody/polyethylene glycol (50 g/L) gave one-third the nonspecific binding and as rapid a separation as with polyethylene glycol (180 g/L) alone. Optimal conditions for separating the immune complex were established, and separation was found to be independent of protein concentrations in urine or serum. This tested assay can detect increasing and ovulatory lutropin concentrations in urine or serum, but with some sacrifice in sensitivity.

125I-radioimmunoassay for unconjugated estriol in serum of pregnant women.

1978 ◽  
Vol 24 (7) ◽  
pp. 1100-1104 ◽  
Author(s):  
R J Liedtke ◽  
J P Greaves ◽  
J D Batjer ◽  
B Busby
Keyword(s):  
Polyethylene Glycol ◽  
Pregnant Women ◽  
Cross Reactivity ◽  
Analytical Recovery ◽  
Reagent Cost ◽  
Procedural Time ◽  
Antibody Complex ◽  
Antigen Antibody ◽  
Entire Procedure ◽  
Unconjugated Estriol

Abstract An 125I-radioimmunoassay is described which measures unconjugated estriol in the serum of pregnant women. Estriol is extracted into ethyl acetate/hexane, an aliquot is evaporated, and the residue is redissolved in phosphate buffer. The sample is incubated with tracer and antibody at 37 degrees C for 15 min and then at 4 degrees C for 1 h and 45 min. The antibody-bound fraction is then precipitated with polyethylene glycol and isolated by centrifugation. Because the antigen-antibody complex is stable in the presence of polyethylene glycol, the separation steps are not influenced by timing. Extraction recovery of 3H-labeled estriol added to a pool of sera from pregnant women averaged 96.7% (SD = 1.3, n = 10). Estriol-supplemented (5, 10, and 20 microgram/liter) serum from men, carried through the entire procedure, showed analytical recovery ranging from 94 to 106%. Structurally analogous steroids normally present in serum of pregnant women exhibit negligible cross reactivity. Day-to-day precision (CV) is 13.3% (3.1 microgram/liter), 6.4% (7.6 microgram/liter), and 5.6% (21.4 microgram/liter) for n = 21. The current reagent cost (about 17 cents per tube), and a total procedural time, including counting, of 5.5 h make this an acceptable assay for routine use.

scholarly journals A Quick & Simple Single-Antibody Immunoassay for the Student Laboratory

2019 ◽  
Vol 81 (3) ◽  
pp. 197-199
Author(s):  
Paul L. Guy
Keyword(s):  
Product Development ◽  
Color Change ◽  
Specific Binding ◽  
Disease Incidence ◽  
Chromogenic Substrate ◽  
Plastic Plate ◽  
Nonspecific Binding ◽  
Student Laboratory ◽  
Antigen Antibody

In this exercise students use a simplified version of an immunoassay that relies on the nonspecific binding of proteins to nitrocellulose followed by the specific binding of antibodies to antigen. This antigen–antibody interaction is detected when a chromogenic substrate, catalyzed by the enzyme conjugated to the antibody, produces a color change. The immunoblot is less expensive and faster to perform than plastic-plate-based assays. I have used this assay for over 10 years in undergraduate courses: in ecology to estimate disease incidence; in botany to explore immunological techniques; and in biotechnology as an exercise in product development.

Influence of Prostaglandin E1 on Slight Proteinuria in Non-Azotaemic Diabetics

1992 ◽  
Vol 20 (1) ◽  
pp. 94-97 ◽  
Author(s):  
S Okada ◽  
K Sato ◽  
T Higuchi ◽  
K Ichiki ◽  
S Tanokuchi ◽  
...  
Keyword(s):  
Insulin Dependent Diabetes Mellitus ◽  
Total Serum Protein ◽  
Dependent Diabetes Mellitus ◽  
Total Serum ◽  
Prostaglandin E ◽  
Albumin Concentration ◽  
Protein Excretion ◽  
Before And After ◽  
Insulin Dependent ◽  
Student’S T

In an investigation into the effect of prostaglandin E1 on proteinuria in nephrotic diabetic nephropathy, five patients were treated with 40 μg prostaglandin E1 administered intravenously over 2 h twice daily for 4 weeks. The following parameters were compared before and after treatment: protein excretion in urine; total serum protein concentration; serum albumin concentration; creatinine clearance; blood urea nitrogen; and serum creatinine content. A further five patients with nephropathy resulting from non-insulin-dependent diabetes mellitus were selected as controls. Analysis of the results using Student's t-test showed no significant change in any of the parameters before and after treatment.

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