Indentity and integraty of 'alfa' and 'beta' - subunit of human thyrotropin prepared by prolonged acetic acid treatment
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Data
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2009
Autores IPEN
CRISTIANE MOREIRA DE CARVALHO
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ANNUAL MEETING OF THE BRAZILIAN BIOCHEMISTRY AND MOLECULAR BIOLOGY SOCIETY, 38.
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Resumo
Alpha- and beta- subunits, prepared by efficiently dissociating, during
16 hours, a recombinant thyrotropin (hTSH) preparation with 0.4 M acetic acid and
isolating them by RP-HPLC, were analysed for what concerns their identity and
integrity. Identity was evaluated by MALDI-TOF mass spectrometry (MALDI-TOF
MS). A relative molecular mass of 14021 and of 15851 was obtained for a-hTSH
and (3-hTSH respectively. These values agree with those obtained by analyzing
the preparation before dissociation, a difference of -1.8% for a and +1.3% for (3
being observed. Integrity of the subunits was evaluated by their capacity of self
reassembling and of restoring the in vivo bioactivity of the hormone. When a-hTSH
and I3-hTSH subunits were incubated together in 0.2 M sodium phosphate buffer,
pH 7.0, at 25°C and under gentle shaking, a complete reassociation occurred after
4 days, forming an heterodimer. In an in vivo mouse bioassay, the T4 levels of the
animals treated with the reassociated hormone were non-significantly different (p>
0.05) from those obtained when the original preparation was administered (2.71±
0.63 pg/dL versus 2.84± 0.23 pg/dL, n=6, respectively). In conclusion, subunits
prepared by prolonged acetic acid treatment maintain their original molecular
mass and can perfectly restore the biological activity of the reassociated
heterodimers.
Como referenciar
ALMEIDA, BEATRIZ E.; CARVALHO, CRISTIANE M.; DAMIANI, RENATA; OLIVEIRA, JOAO E.; BARTOLINI, PAOLO; RIBELA, MARIA T.C.P. Indentity and integraty of 'alfa' and 'beta' - subunit of human thyrotropin prepared by prolonged acetic acid treatment. In: ANNUAL MEETING OF THE BRAZILIAN BIOCHEMISTRY AND MOLECULAR BIOLOGY SOCIETY, 38., May 16-19, 2009, Aguas de Lindoia, SP. Abstract... Disponível em: http://repositorio.ipen.br/handle/123456789/19346. Acesso em: 21 Jul 2026.
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