Measurement of cAMP accumulation in Chinese hamster ovary cells transfected with the recombinant human TSH receptor (CHO-R)

a new bioassay for human thyrotropin

L. Persani, M. Tonacchera, P. Beck-Peccoz, P. Vitti, C. Mammoli, L. Chiovato, R. Elisei, G. Faglia, M. Ludgate, G. Vassart, Aldo Pinchera

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Circulating TSH bioactivity may vary in several clinical and experimental conditions. Since the reliability of the current methods for the measurement of TSH bioactivity is limited, a new bioassay based on cAMP accumulation in Chinese Hamster Ovary cells transfected with recombinant human TSH receptor (CHO-R) was set up. The sensitivity was 0.3±0.1, 0.4±0.1 and 0.01±0.01 μg/L for TSH IRP 80/558, recombinant human TSH and bovine TSH, respectively. Standard curves were parallel, and the intra- and inter-assay coefficients of variation were 13±1.1% and 22±1.9%, respectively. LH, FSH, CG and TSH subunits did not stimulate cAMP accumulation up to high concentrations. Circulating TSH was partially purified by immunoaffinity separation and concentrated before being bioassayed. However, plain sera with high TSH levels, such as those from primary hypothyroid patients (PH), could be directly tested in CHO-R bioassay, provided that sera were added at concentrations lower than 10%. TSH from 6 normal subjects had biological to immunological ratio (B/l) ranging from 0.6 to 2.1 (mean±SD=1.4±0.5). TSH from 6 patients with PH showed bioactivity significantly lower than in normals (B/l=0.6±0.3; p

Original languageEnglish
Pages (from-to)511-519
Number of pages9
JournalJournal of Endocrinological Investigation
Volume16
Issue number7
DOIs
Publication statusPublished - 1993

Fingerprint

Thyrotropin Alfa
Thyrotropin Receptors
Thyrotropin
Cricetulus
Biological Assay
Ovary
Serum

Keywords

  • central hypothyroidism
  • CHO cells
  • FRTL-5
  • primary hypothyroidism
  • Thyrotropin
  • TRH
  • TSH bioactivity
  • TSH receptor

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Measurement of cAMP accumulation in Chinese hamster ovary cells transfected with the recombinant human TSH receptor (CHO-R) : a new bioassay for human thyrotropin. / Persani, L.; Tonacchera, M.; Beck-Peccoz, P.; Vitti, P.; Mammoli, C.; Chiovato, L.; Elisei, R.; Faglia, G.; Ludgate, M.; Vassart, G.; Pinchera, Aldo.

In: Journal of Endocrinological Investigation, Vol. 16, No. 7, 1993, p. 511-519.

Research output: Contribution to journalArticle

Persani, L. ; Tonacchera, M. ; Beck-Peccoz, P. ; Vitti, P. ; Mammoli, C. ; Chiovato, L. ; Elisei, R. ; Faglia, G. ; Ludgate, M. ; Vassart, G. ; Pinchera, Aldo. / Measurement of cAMP accumulation in Chinese hamster ovary cells transfected with the recombinant human TSH receptor (CHO-R) : a new bioassay for human thyrotropin. In: Journal of Endocrinological Investigation. 1993 ; Vol. 16, No. 7. pp. 511-519.
@article{e58dbae4e9dd4a07ba0b4d2d3e8f7d6a,
title = "Measurement of cAMP accumulation in Chinese hamster ovary cells transfected with the recombinant human TSH receptor (CHO-R): a new bioassay for human thyrotropin",
abstract = "Circulating TSH bioactivity may vary in several clinical and experimental conditions. Since the reliability of the current methods for the measurement of TSH bioactivity is limited, a new bioassay based on cAMP accumulation in Chinese Hamster Ovary cells transfected with recombinant human TSH receptor (CHO-R) was set up. The sensitivity was 0.3±0.1, 0.4±0.1 and 0.01±0.01 μg/L for TSH IRP 80/558, recombinant human TSH and bovine TSH, respectively. Standard curves were parallel, and the intra- and inter-assay coefficients of variation were 13±1.1{\%} and 22±1.9{\%}, respectively. LH, FSH, CG and TSH subunits did not stimulate cAMP accumulation up to high concentrations. Circulating TSH was partially purified by immunoaffinity separation and concentrated before being bioassayed. However, plain sera with high TSH levels, such as those from primary hypothyroid patients (PH), could be directly tested in CHO-R bioassay, provided that sera were added at concentrations lower than 10{\%}. TSH from 6 normal subjects had biological to immunological ratio (B/l) ranging from 0.6 to 2.1 (mean±SD=1.4±0.5). TSH from 6 patients with PH showed bioactivity significantly lower than in normals (B/l=0.6±0.3; p",
keywords = "central hypothyroidism, CHO cells, FRTL-5, primary hypothyroidism, Thyrotropin, TRH, TSH bioactivity, TSH receptor",
author = "L. Persani and M. Tonacchera and P. Beck-Peccoz and P. Vitti and C. Mammoli and L. Chiovato and R. Elisei and G. Faglia and M. Ludgate and G. Vassart and Aldo Pinchera",
year = "1993",
doi = "10.1007/BF03348894",
language = "English",
volume = "16",
pages = "511--519",
journal = "Journal of Endocrinological Investigation",
issn = "0391-4097",
publisher = "Springer International Publishing",
number = "7",

}

TY - JOUR

T1 - Measurement of cAMP accumulation in Chinese hamster ovary cells transfected with the recombinant human TSH receptor (CHO-R)

T2 - a new bioassay for human thyrotropin

AU - Persani, L.

AU - Tonacchera, M.

AU - Beck-Peccoz, P.

AU - Vitti, P.

AU - Mammoli, C.

AU - Chiovato, L.

AU - Elisei, R.

AU - Faglia, G.

AU - Ludgate, M.

AU - Vassart, G.

AU - Pinchera, Aldo

PY - 1993

Y1 - 1993

N2 - Circulating TSH bioactivity may vary in several clinical and experimental conditions. Since the reliability of the current methods for the measurement of TSH bioactivity is limited, a new bioassay based on cAMP accumulation in Chinese Hamster Ovary cells transfected with recombinant human TSH receptor (CHO-R) was set up. The sensitivity was 0.3±0.1, 0.4±0.1 and 0.01±0.01 μg/L for TSH IRP 80/558, recombinant human TSH and bovine TSH, respectively. Standard curves were parallel, and the intra- and inter-assay coefficients of variation were 13±1.1% and 22±1.9%, respectively. LH, FSH, CG and TSH subunits did not stimulate cAMP accumulation up to high concentrations. Circulating TSH was partially purified by immunoaffinity separation and concentrated before being bioassayed. However, plain sera with high TSH levels, such as those from primary hypothyroid patients (PH), could be directly tested in CHO-R bioassay, provided that sera were added at concentrations lower than 10%. TSH from 6 normal subjects had biological to immunological ratio (B/l) ranging from 0.6 to 2.1 (mean±SD=1.4±0.5). TSH from 6 patients with PH showed bioactivity significantly lower than in normals (B/l=0.6±0.3; p

AB - Circulating TSH bioactivity may vary in several clinical and experimental conditions. Since the reliability of the current methods for the measurement of TSH bioactivity is limited, a new bioassay based on cAMP accumulation in Chinese Hamster Ovary cells transfected with recombinant human TSH receptor (CHO-R) was set up. The sensitivity was 0.3±0.1, 0.4±0.1 and 0.01±0.01 μg/L for TSH IRP 80/558, recombinant human TSH and bovine TSH, respectively. Standard curves were parallel, and the intra- and inter-assay coefficients of variation were 13±1.1% and 22±1.9%, respectively. LH, FSH, CG and TSH subunits did not stimulate cAMP accumulation up to high concentrations. Circulating TSH was partially purified by immunoaffinity separation and concentrated before being bioassayed. However, plain sera with high TSH levels, such as those from primary hypothyroid patients (PH), could be directly tested in CHO-R bioassay, provided that sera were added at concentrations lower than 10%. TSH from 6 normal subjects had biological to immunological ratio (B/l) ranging from 0.6 to 2.1 (mean±SD=1.4±0.5). TSH from 6 patients with PH showed bioactivity significantly lower than in normals (B/l=0.6±0.3; p

KW - central hypothyroidism

KW - CHO cells

KW - FRTL-5

KW - primary hypothyroidism

KW - Thyrotropin

KW - TRH

KW - TSH bioactivity

KW - TSH receptor

UR - http://www.scopus.com/inward/record.url?scp=0027234570&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027234570&partnerID=8YFLogxK

U2 - 10.1007/BF03348894

DO - 10.1007/BF03348894

M3 - Article

VL - 16

SP - 511

EP - 519

JO - Journal of Endocrinological Investigation

JF - Journal of Endocrinological Investigation

SN - 0391-4097

IS - 7

ER -