An accurate determination of human growth hormone content in different pituitary extracts, using a radioimmunoassay with polyacrylamide gel electrophoresis as a bound-free separation system

Bartolini, Paolo; Assis, Luís Marques de; Schwarz, Irene; Pieroni, R.R.

doi:10.1016/0009-8981(77)90481-8

Cited by 10 publications

(2 citation statements)

References 18 publications

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“…Serum growth hormone was determined by radioimmunoassay using human growth hormone prepared in this laboratory (Bartolini, Assis, Schwarz and Pieroni 1977). The separation of bound and free hormone was accomplished with PEG (Desbuquois and Aurbach 1971).…”

Section: Methodsmentioning

confidence: 99%

The “So-Called” Somatomedin B and Growth Hormone Measured by Radioimmunoassay in Normal and Cirrhotic Individuals

Fh¹,

Oz²,

Aa³

et al. 1984

Horm Metab Res

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Serum somatomedin B (SmB) levels in cirrhotic individuals, 3.3 +/- 1.5 mg/l, were strikingly lower (P less than 0.001) than in normal subjects, 9.0 +/- 1.7 mg/l. SmB levels were clearly related to the levels of alpha 2-globulins in the cirrhotics (r = + 0.8, P less than 0.002). Serum SmB and growth hormone correlated negatively in a group of normal and cirrhotic individuals (r = -0.67, P less than 0.001). Direct measurements of serum SmB failed to reveal differences between hepatic, renal and peripheral veins. These findings suggest that: 1) SmB is produced by liver and/or normal liver function plays an important role in maintaining normal serum SmB levels; 2) SmB carrier proteins are reduced in the cirrhotics and 3) SmB is part of a negative feed-back system involving growth hormone.

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Section: Methodsmentioning

confidence: 99%

The “So-Called” Somatomedin B and Growth Hormone Measured by Radioimmunoassay in Normal and Cirrhotic Individuals

Fh¹,

Oz²,

Aa³

et al. 1984

Horm Metab Res

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show abstract

“…Aliquots after one, four, nine, twenty and twenty-one months of storage were iodinated and purified on QAE-Sephadex A 25 (Jørgensen and Larsen 1972). The main peak-related fraction, from the tracer purification, and one sample of the unlabeled hormone were submitted to PAGE analysis (Bartolini, Assis, Schwarz and Pieroni 1977). The radioactive gel was cut into segments (0.7 cm) and counted, while the cold gel was stained in Coomassie brilliant blue.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Glucagon Stability on Storage for Radioiodination Purposes

Borghi¹,

Wajchenberg²

1982

Horm Metab Res

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Ultrasensitive in vivo bioassay detects bioactive human growth hormone in transduced primary human keratinocytes

Bellini

Mathor

Luca

et al. 1998

J Endocrinol Invest

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An improved in vivo body weight gain bioassay for the potency determination of human growth hormone (hGH) has been set up in "little" mice (lit/lit), a mutant derived from the C57BL/6J strain. This improved assay now has a detection limit of the order of 0.05 micrograms/mouse/day, which corresponds to a sensitivity about 20-fold higher than that of the most sensitive in vivo assay reported up to now: the tibia test in hypophysectomized rats or mice. This sensitivity was achieved mainly by introduction of a careful pre-assay selection and of a three injections per day schedule. The utilization of these conditions in a 2x2 factorial assay design allowed the potency determination of recombinant DNA-derived hGH (rec-hGH) in bacterial extracts with acceptable accuracy and precision, together with the greatest economy of material, only 0.24 mg of unknown and standard hormone preparation being sufficient for an entire 10-animal assay. This contrasts to a minimum of 2.7 mg that are necessary for an economical assay in hypophysectomized rats. The same assay procedure was also used to demonstrate the in vivo bioactivity of hGH secreted into a culture medium from transduced human primary keratinocytes. The growth curve constructed with n = 8 little mice presented a highly significant correlation (r = 0.939, p < 0.001) and a slope = 0.016 g/mouse/day. It was thus possible to prove, for the first time, the in vivo bioactivity of rec-hGH secreted by transplantable human epidermal cells, utilized as an experimental model for somatic gene therapy.

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An accurate determination of human growth hormone content in different pituitary extracts, using a radioimmunoassay with polyacrylamide gel electrophoresis as a bound-free separation system

Cited by 10 publications

References 18 publications

The “So-Called” Somatomedin B and Growth Hormone Measured by Radioimmunoassay in Normal and Cirrhotic Individuals

The “So-Called” Somatomedin B and Growth Hormone Measured by Radioimmunoassay in Normal and Cirrhotic Individuals

Evaluation of Glucagon Stability on Storage for Radioiodination Purposes

Ultrasensitive in vivo bioassay detects bioactive human growth hormone in transduced primary human keratinocytes

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