IMMUNOCHEMICAL STUDIES ON THE HETEROGENEITY OF HUMAN GROWTH HORMONE (HGH)

1969 ◽  
Vol 60 (1) ◽  
pp. 101-111 ◽  
Author(s):  
W. Rohde ◽  
G. Dörner
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Extraction Procedure ◽  
Gel Filtration ◽  
Human Growth ◽  
Double Diffusion ◽  
Active Components ◽  
Agar Gel ◽  
Human Pituitary ◽  
Starch Gel

ABSTRACT The specific antigenic patterns of human growth hormone in various human pituitary extracts were determined by analysis in gel filtration, in starch gel and agar gel immunoelectrophoresis. The analysis of pituitary extracts by gel filtration on Sephadex G-200 indicates that HGH complexes with different molecular weights were present in all tested pituitary extracts. The rate of formation of high molecular HGH was dependent on the extraction procedure used. Altogether seven immunologically active components of HGH were detected by analysis of crude pituitary extracts in starch gel immunoelectrophoresis. The number and the rate of formed HGH components were dependent on the solvent used for the extraction of pituitary homogenates and on the sequence of extraction procedures. The electrophoretical patterns obtained were reproducible. The electrophoretical behaviour of HGH of various pituitary extracts in agar gel also suggested the presence of multiple components. Three different positions were observed between the α1- and β1-globulins. The analysis of pituitary extracts in the two dimensional double diffusion technique in agar gel using an absorbed anti-HGH-serum gave evidence of the antigenic homogeneity of HGH in the different preparations. It is concluded that the electrophoretic heterogeneity of pituitary growth hormone preparations is already determined by the first step of the extraction.

IMMUNOCHEMICAL STUDIES OF HUMAN GROWTH HORMONE, OVINE PROLACTIN, BOVINE GROWTH HORMONE AND A TRYPTIC DIGEST OF BOVINE GROWTH HORMONE

1969 ◽  
Vol 44 (4) ◽  
pp. 517-NP ◽  
Author(s):  
KALYAN SUNDARAM ◽  
M. SONENBERG
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Deae Cellulose ◽  
Human Growth ◽  
Double Diffusion ◽  
Bovine Growth Hormone ◽  
Agar Gel ◽  
Tryptic Digest ◽  
Immunological Reactions ◽  
Ovine Prolactin

SUMMARY Agar gel double diffusion and immunoelectrophoresis of human growth hormone (HGH), ovine prolactin, bovine growth hormone (BGH) and a tryptic digest of BGH (TBGH) were performed. Antisera to BGH, TBGH, HGH and ovine prolactin were used for the tests. Both BGH and TBGH were found to contain material that precipitated with antiserum to ovine prolactin. Immunological analysis of various fractions of BGH and TBGH separated by chromatography on DEAE-cellulose showed that BGH and TBGH were eluted before prolactin. No differences between BGH and TBGH in their immunological reactions were found. HGH did not cross-react with BGH, TBGH or ovine prolactin.

IMMUNOCHEMICAL STUDIES OF HUMAN PITUITARY GROWTH HORMONE

1962 ◽  
Vol 40 (2) ◽  
pp. 311-320 ◽  
Author(s):  
Zvi Laron ◽  
Sara Assa
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Haemagglutination Inhibition ◽  
Rabbit Antiserum ◽  
Human Growth ◽  
Human Albumin ◽  
Agar Gel ◽  
Human Pituitary ◽  
Pituitary Glands ◽  
Fraction I

ABSTRACT Human growth hormone (HGH) prepared from human pituitary glands by the method of Raben (1959) showed two components on agar gel electrophoresis. Fraction I corresponded to a slow alpha-globulin, and fraction II had a mobility between alpha and beta-globulin. On agar gel immunoelectrophoresis using HGH and rabbit antiserum to HGH, these two components showed immunological identity. By using agar gel immunoelectrophoresis and the haemagglutination technique HGH preparations of both Raben and Li type were shown to be contaminated with human albumin. Antiserum to human growth hormone containing also antibodies against human albumin was purified by adsorption with human albumin. It is postulated that part of the difficulties encountered in the use of the haemagglutination-inhibition technique to determine growth hormone concentration in serum, is due to impurities in the human growth hormone preparations and antisera to these preparations in use.

scholarly journals Isolation and properties of the electrophoretic components of human growth hormone by Sephadex-gel filtration and preparative polyacrylamide-gel electrophoresis

1966 ◽  
Vol 100 (3) ◽  
pp. 711-717 ◽  
Author(s):  
BB Saxena ◽  
PH Henneman
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Polyacrylamide Gel Electrophoresis ◽  
Group Analysis ◽  
Gel Filtration ◽  
Human Growth ◽  
Circular Disk ◽  
Biologically Active ◽  
Human Pituitary ◽  
Three Components

1. Human growth hormone was prepared from acetone-dried residues after extraction of gonadotrophins from pituitary glands. 2. Crude growth hormone was purified by gel filtration on Sephadex, resulting in a product that is soluble in water or 0.5% sodium chloride. It is painless on injection and shows a twofold increase in biological potency. Aggregation of growth hormone on Sephadex columns can be avoided by the addition of urea (6m) and EDTA (1mm) to the buffer. 3. Growth hormone appeared as a single component from Sephadex and ion-exchange columns and sedimented as a single boundary in the ultracentrifuge. In the circular disk electrophoresis, however, the growth hormone showed one faster and two slower-moving anionic components. 4. These components were isolated by preparative electrophoresis on polyacrylamide columns. The purified growth hormone and its three components sedimented as single boundaries with coefficients 2.62, 2.66, 2.66 and 2.83s respectively. 5. Amino acid analyses of the purified growth hormone and its components were closely related. End-group analysis of purified growth hormone and its components showed only phenylalanine at both N- and C-terminals. 6. The purified growth hormone and its components were essentially free of other pituitary hormones, but contained significant prolactin activity. The biochemical similarities among the electrophoretic components of human growth hormone and the presence of the same three components in the growth hormone prepared from a single human pituitary gland suggest polymorphism of a biologically active protein molecule.

EFFECT OF HEAT ON THE IMMUNOLOGICAL PROPERTIES OF HUMAN, BOVINE AND OVINE GROWTH HORMONE

1964 ◽  
Vol 46 (3) ◽  
pp. 465-472 ◽  
Author(s):  
Zvi Laron ◽  
Ariana Yed-Lekach ◽  
Sara Assa ◽  
Avivah Kowadlo-Silbergeld
Keyword(s):  
Heat Treatment ◽  
Growth Hormone ◽  
Electrophoretic Mobility ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Precipitation Reaction ◽  
Agar Gel ◽  
Immunological Properties ◽  
Haemagglutination Test ◽  
Precipitation Reactions

ABSTRACT Human, bovine and sheep (ovine) growth hormone (HGH, BGH and SGH) were heated in solution at temperatures between 60 and 100 °C. The electrophoretic mobility and immunological properties, such as precipitation reactions in agar gel and haemagglutination with antiserum to untreated hormone, were studied at different degrees of heating. It was found that heat progressively reduced the immunological properties of the growth hormone; however, human growth hormone was more resistant to heat treatment than the bovine and sheep growth hormone. HGH retained precipitation properties when heated at 100° C up to 30 minutes, and reacted in the haemagglutination test when heated at 100° C for less than 60 minutes. BGH and SGH clotted at 100° C. The precipitation reaction with antiserum to BGH disappeared when BGH or SGH was heated at 70° C for more than 10 minutes. Only a weak haemagglutination reaction was retained when BGH or SGH was heated at 80° C for 15 minutes.

Radioimmunoassay of Human Growth Hormone

1968 ◽  
Vol 14 (2) ◽  
pp. 145-155 ◽  
Author(s):  
Mechthilde Knoller ◽  
M U Tsao ◽  
George H Lowrey
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Gel Filtration ◽  
Human Growth ◽  
Reproducible Method

Abstract Detailed methods for: (1) 131I iodination of human growth hormone (HGH); (2) purification of labeled HGH (HGH-131I; and (3) radioimmunoassay of HGH are given. Bio-Gel filtration is introduced as a rapid and reproducible method for purifying HGH-131I which has been obtained by a modification of the method of Greenwood et al. (1). Highly purified HGH-131I with a bindability of 96% or more is usually achieved.

STUDIES ON A MONKEY PLACENTAL PROTEIN WITH IMMUNOCHEMICAL SIMILARITY TO HUMAN GROWTH HORMONE AND HUMAN CHORIONIC SOMATOMAMMOTROPHIN

1970 ◽  
Vol 63 (4) ◽  
pp. 736-746 ◽  
Author(s):  
David B. Grant ◽  
Selna L. Kaplan ◽  
Melvin M. Grumbach
Keyword(s):  
Growth Hormone ◽  
Electrophoretic Mobility ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Double Diffusion ◽  
Molecular Weights ◽  
Diffusion Studies ◽  
Placental Protein ◽  
Two Peaks ◽  
Dilution Curves

ABSTRACT The properties of a monkey placental protein (monkey chorionic somatomammotrophin; MCS) which reacts with antisera directed against human growth hormone (HGH) and human chorionic somatomammotrophin (HCS) have been studied. Following application of monkey placental extracts to Sephadex G-100 and elution with 0.1 m NH4HCO3, two peaks of MCS, with molecular weights of approximately 25 000 and 50 000 were obtained. The electrophoretic mobility of MCS in acrylamide at pH 8.2–8.4 was similar to but less anodal than that of HGH or HCS. The results of immunoelectrophoretic and Ouchterlony double-diffusion studies indicated that MCS is physicochemically and immunochemically distinguishable from HGH and HCS. When tested in a radioimmunoassay for HGH, MCS gave dilution curves intermediate between those for HGH and HCS. In contrast, MCS and HGH gave similar curves in a radioimmunoassay for HCS. MCS, HCS and HGH had very similar effects on the reactions between 131I-HGH and anti-HCS or 131I-HCS and anti-HGH serum. Radioimmunoassay systems for HCS were unsuitable for estimating MCS; however, the above mentioned hybrid assay systems provide a means for approximate quantitation of MCS.

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