scholarly journals Immunoassay of serum polypeptide hormones by using 125I-labelled anti(-immunoglobulin G) antibodies

1975 ◽  
Vol 145 (3) ◽  
pp. 607-616 ◽  
Author(s):  
P Beck ◽  
H Nicholas
Keyword(s):  
Growth Hormone ◽  
Parathyroid Hormone ◽  
Guinea Pig ◽  
Human Growth Hormone ◽  
Immunoglobulin G ◽  
Human Growth ◽  
Bovine Insulin ◽  
Polypeptide Hormone ◽  
Polypeptide Hormones ◽  
Bovine Parathyroid Hormone

1. A technique for indirectly labelling antibodies to polypeptide hormones, by combining them with radioactively labelled anti-(immunoglobulin G) is described. (a) 125I-labelled anti-(rabbit immunoglobulin G) and anti-(guinea-pig immunoglobulin G) antibodies with high specific radioactivity were prepared after purification of the antibodies on immunoadsorbents containing the respective antigens. (b) Rabbit immunoglobulin G antibodies to human growth hormone, porcine glucagon and guinea-pig immunoglobulin G antibodies to bovine insulin and bovine parathyroid hormone were combined with immunoadsorbents containing the respective polypeptide hormone antigen. (c) The immunoglobulin G antibodies to the polypeptide hormones were reacted with 125-I-labelled anti-(immunoglobulin G) antibodies directed against the appropriate species of immunoglobulin G,and the anti-hormone antibodies were combined with the hormone-containing immunoadsorbent. (d) 125I-labelled anti-(immunoglobulin G) antibodies and anti-hormone antibodies were simultaneously eluted from the hormone-containing immunoadsorbent by dilute HCl, pH 2.0. After elution the anti-(immunoglobulin G) antibodies and antihormone antibodies were allowed to recombine at pH 8.0 and 4 degrees C. 2. The resultant immunoglobulin G-anti-immunoglobulin G complex was used in immunoradiometric (labelled antibody) and two-site assays of the respective polypeptide hormone. 3. By using these immunoassays, concentrations down to 90pg of human growth hormone/ml, 100 pg of bovine insulin/ml, 80 pg of bovine parathyroid hormone/ml and 150 pg of glucagon/ml were readily detected. Assays of human plasma for growth hormone and insulin by these methods showed good agreement with results obtained by using a directly 125I-labelled anti-hormone antibody in an immunoradiometric assay of human growth hormone or by radioimmunoassay of human insulin. 4. The method described allows immunoradiometric or two-site assays to be performed starting with as little as 450 ng of polypeptide hormone-antibody protein. An additional advantage of the method is that a single iodination of the readily available antibodies to immunoglobulin G allows the establishemnt of several polypeptide hormone assays

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.

GROWTH HORMONE AND GROWTH

PEDIATRICS ◽  
1966 ◽  
Vol 37 (2) ◽  
pp. 245-248
Author(s):  
MELVIN M. GRUMBACH
Keyword(s):  
Growth Hormone ◽  
Pituitary Gland ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Analytical Technique ◽  
The Past ◽  
New Information ◽  
Polypeptide Hormones ◽  
The Subject ◽  
Methods Of Isolation

THE RECENT cascade of knowledge of the chemistry and physiology of the protein and polypeptide hormones of the pituitary gland has been a consequence of steady progress over the past 20 years in methods of isolation, purification, and assay of these hormones. These development, while providing new insights into old problems, have raised a host of challenging and unanticipated questions. Of especial interest to students of growth is the considerable new information on growth hormone, the subject of a recent and perceptive review by Allen Root. New analytical technique utilizing immunochemical methods and radioiodinated hormone have provided specific, sensitive, and precise redioimmunoassay procedures for the measurement of the minute quanitity of human growth hormone in blood.

LIPOLYTIC ACTIVITY OF ENZYME DIGESTED HUMAN GROWTH HORMONE

1965 ◽  
Vol 49 (3_Suppl) ◽  
pp. S143
Author(s):  
Zvi Laron ◽  
Avivah Kowadlo-Silbergeld
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Lipolytic Activity ◽  
Human Growth
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