PMID- 8563133
OWN - NLM
STAT- MEDLINE
DCOM- 19960307
LR  - 20190512
IS  - 0959-6658 (Print)
IS  - 0959-6658 (Linking)
VI  - 5
IP  - 5
DP  - 1995 Jul
TI  - Biological and immunochemical characterization of recombinant human thyrotrophin.
PG  - 473-81
AB  - Recombinant human thyroid-stimulating hormone (recTSH) has recently been 
      engineered to detect metastatic lesions in patients operated on for thyroid 
      cancer. In this report, we have compared the microheterogeneity, carbohydrate 
      (CHO) content, mitogenic potency and immunoreactivity of the biotechnology 
      product to those of human TSH of pituitary origin (pitTSH). Compositional 
      analysis revealed that recombinant (rec) TSH produced in Chinese hamster ovary 
      cells was overglycosylated compared with the native hormone (21 and 14%, 
      respectively) with a higher amount of sialic acid and lack of 
      N-acetylgalactosamine. Electrofocusing followed by immunoblotting resolved recTSH 
      into six glycoforms with pIs ranging from 6.0 to 8.6, which were converted to a 
      major species of pI 8.9 by sialidase treatment. pitTSH contained five main 
      isoforms of pI 6.5-8.2 distinct from those of recTSH and partially resistant to 
      sialidase. Binding activity of both human TSHs to porcine thyroid membrane 
      receptors was found to be similar, but recTSH appeared to be 20% active compared 
      to pitTSH in eliciting cAMP production and cell growth in rat FRTL-5 cells. 
      Immunoreactivity of the recombinant hormone was investigated using polyclonal and 
      monoclonal antibodies raised against the native hormone or synthetic peptide 
      sequences of its subunits. While rec- and pitTSH were recognized to a similar 
      extent by anti-protein antibodies, they exhibited a different binding pattern to 
      antipeptide antibodies. Serial dilution of anti-alpha 1-25, anti-alpha 26-51, 
      anti-beta 96-112 antisera bound recTSH to a greater extent than pitTSH, while 
      anti-beta 31-51 and anti-beta 53-76 displayed similar recognition toward both 
      preparations.(ABSTRACT TRUNCATED AT 250 WORDS)
FAU - Canonne, C
AU  - Canonne C
AD  - Laboratoire d'Immunochimie des Hormones Glycoproteiques and U 270 INSERM, Faculte 
      de Medecine Nord, Marseille, France.
FAU - Papandreou, M J
AU  - Papandreou MJ
FAU - Medri, G
AU  - Medri G
FAU - Verrier, B
AU  - Verrier B
FAU - Ronin, C
AU  - Ronin C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Glycobiology
JT  - Glycobiology
JID - 9104124
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Epitopes)
RN  - 0 (Receptors, Thyrotropin)
RN  - 0 (Recombinant Proteins)
RN  - 9002-71-5 (Thyrotropin)
RN  - EC 3.2.1.18 (Neuraminidase)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/metabolism
MH  - Binding, Competitive
MH  - CHO Cells/metabolism
MH  - Carbohydrate Metabolism
MH  - Cell Division/drug effects
MH  - Cricetinae
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Epitopes/metabolism
MH  - Glycosylation
MH  - Humans
MH  - Immunochemistry
MH  - Neuraminidase/pharmacology
MH  - Pituitary Gland/metabolism
MH  - Protein Engineering
MH  - Rats
MH  - Receptors, Thyrotropin/metabolism
MH  - Recombinant Proteins/chemistry/drug effects/genetics/immunology/metabolism
MH  - Swine
MH  - Thyrotropin/*chemistry/drug effects/genetics/immunology/metabolism
EDAT- 1995/07/01 00:00
MHDA- 1995/07/01 00:01
CRDT- 1995/07/01 00:00
PHST- 1995/07/01 00:00 [pubmed]
PHST- 1995/07/01 00:01 [medline]
PHST- 1995/07/01 00:00 [entrez]
AID - 10.1093/glycob/5.5.473 [doi]
PST - ppublish
SO  - Glycobiology. 1995 Jul;5(5):473-81. doi: 10.1093/glycob/5.5.473.