RADIOIMMUNOASSAY OF HUMAN GROWTH HORMONE USING THE DOUBLE ANTIBODY TECHNIQUE

1968 ◽  
Vol 57 (4) ◽  
pp. 549-556 ◽  
Author(s):  
F. Melani ◽  
U. Gröschel-Stewart ◽  
J. Lawecki
Keyword(s):  
Growth Hormone ◽  
Human Growth ◽  
Biologically Active ◽  
List Type ◽  
Purification Step ◽  
Antibody Technique ◽  
Double Antibody ◽  
Starch Gel ◽  
Radio Immunoassay

ABSTRACT The method of Roos et al. (1963) for the preparation of HGH yields a highly purified biologically active hormone preparation suitable for clinical purposes and, after one additional purification step (chromatography on Sephadex G-200 according to Hunter (1965), for radioiodination. In the double antibody technique described, the labelled hormone can be used for radio-immunoassay without starch-gel or Sephadex purification. The method is simple and sensitive enough to permit the determination of HGH in serum.

IMMUNOREACTIVE GROWTH HORMONE IN HUMAN URINE

1972 ◽  
Vol 71 (4) ◽  
pp. 665-676 ◽  
Author(s):  
Kristian F. Hanssen
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Linear Relationship ◽  
Human Urine ◽  
Gel Filtration ◽  
Human Growth ◽  
Normal Subjects ◽  
Normal Human ◽  
Radio Immunoassay

ABSTRACT By using a double antibody radio-immunoassay (pre-precipitation technique) for the determination of immunoreactive human growth hormone (IRHGH) in normal human urine concentrated by dialysis and lyophilization, a factor was revealed that displaces 125I-HGH from HGH antibodies. This displacement was neither due to salts nor to glucose; it is suggested that it is due to IRHGH in the urine. A linear relationship between dilution of urine and the measured IRHGH concentration was obtained. Recovery of exogenous HGH was between 70–105%. The recovery of IRHGH from different volumes of urine following dialysis and lyophilization was between 97–110%. Plasma IRHGH and urinary IRHGH was measured simultaneously after HGH injection in a normal subject. A correlation was shown between plasma IRHGH and urinary IRHGH. In 9 normal subjects, the urinary IRHGH ranged from 28–53 ng/24 h. The excretion of urinary IRHGH was increased in acromegaly and was diminished in some, but not in all patients with adult hypopituitarism. The urinary IRHGH was further studied by gel filtration. It was recovered in one peak corresponding to a molecular weight of approximately 20 000 – 30 000. However, in the present work it was not clarified whether the urinary IRHGH represents pituitary HGH excreted in the urine or a metabolite of high molecular weight with retained immunological properties.

DETERMINATION OF HUMAN GROWTH HORMONE (HGH) IN PLASMA BY A DOUBLE ANTIBODY RADIOIMMUNOASSAY

1966 ◽  
Vol 53 (1) ◽  
pp. 101-120 ◽  
Author(s):  
E. Cerasi ◽  
L. Della Casa ◽  
R. Luft ◽  
A. Roovete
Keyword(s):  
Present Method ◽  
Human Growth ◽  
Antibody Technique ◽  
Double Antibody ◽  
Insulin In Plasma ◽  
Assay Procedure ◽  
Absolute Measure ◽  
Assay Methods ◽  
Per Se

ABSTRACT The double antibody technique of Hales & Randle (1963) for determination of insulin in plasma was developed for the assay of HGH in plasma. The precision and reproducibility of the method as well as the recovery of HGH added to plasma were highly satisfactory. Increased HGH values were obtained in acromegaly and during insulin hypoglycaemia, while decreased values were observed during glucose infusion. Successful treatment of acromegaly was accompanied by the return of plasma HGH levels toward normal. The plasma HGH values obtained with the present method cannot be considered as reflecting the absolute amounts of HGH, since several factors in the assay procedure, mainly the incubation time and the concentration of HGH antibodies used, markedly influenced the values obtained. It is questioned whether the radioimmunological assay methods for HGH per se give an absolute measure of this hormone in plasma.

Optimization of Radioimmunoassay for Human Growth Hormone by the Charcoal-Dextran Technique

1970 ◽  
Vol 16 (10) ◽  
pp. 845-848 ◽  
Author(s):  
Joseph C Meek ◽  
Mary M Stoskopf ◽  
Robert E Bolinger
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Guinea Pig ◽  
Human Growth Hormone ◽  
Serum Proteins ◽  
Human Growth ◽  
Molecular Fragments ◽  
Antibody Technique ◽  
Double Antibody

Abstract The effect of serum on the radioimmunoassay for human growth hormone was tested by use of the double-antibody technique and the charcoal-coated dextran technique. With the double-antibody technique, serum effects a falsely high reading for unknowns, while with the charcoal-dextran technique it causes falsely low values to be obtained. Part of the anomaly with the charcoal-dextran technique is the result of adsorption of small molecular fragments of radiolabeled hormone onto serum proteins of higher molecular weight. These fragments can be eliminated by frequent repurification of the labeled hormone. Another part is not related to purity of the label but is corrected by subtracting values for serum blanks that contain no antibody. The use of antiserum to human growth hormone prepared in the goat is more sensitive than that prepared in the guinea pig.

High-level production and one-step purification of biologically active human growth hormone in Escherichia coli

Gene ◽  
1995 ◽  
Vol 165 (2) ◽  
pp. 303-306 ◽  
Author(s):  
Reema Mukhija ◽  
Prithy Rupa ◽  
Devika Pillai ◽  
Lalit C. Garg
Keyword(s):  
Escherichia Coli ◽  
Growth Hormone ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Biologically Active ◽  
One Step ◽  
High Level ◽  
Level Production

scholarly journals ELISA for Determination of Human Growth Hormone: Recognition of Helix 4 Epitopes

2004 ◽  
Vol 2004 (3) ◽  
pp. 143-149 ◽  
Author(s):  
Juliana F. Moura ◽  
Luiz DeLacerda ◽  
Romolo Sandrini ◽  
Fernanda M. Borba ◽  
Denise N. Castelo ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Synthetic Peptide ◽  
Human Growth ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunoaffinity Column ◽  
Receptor Dimerization ◽  
Human Prolactin

Human growth hormone (hGH) signal transduction initiates with a receptor dimerization in which one molecule binds to the receptor through sites 1 and 2. A sandwich enzyme-linked immunosorbent assay was developed for quantifying hGH molecules that present helix 4 from binding site 1. For this, horse anti-rhGH antibodies were eluted by an immunoaffinity column constituted by sepharose-rhGH. These antibodies were purified through a second column with synthetic peptide correspondent tohGH helix 4, immobilized to sepharose, and used as capture antibodies. Those that did not recognize synthetic peptide were used as a marker antibody. The working range was of 1.95 to 31.25 ng/mL of hGH. The intra-assay coefficient of variation (CV) was between 4.53% and 6.33%, while the interassay CV was between 6.00% and 8.27%. The recovery range was between 96.0% to 103.8%. There was no cross-reactivity with human prolactin. These features show that our assay is an efficient method for the determination of hGH.

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