High production and optimization of the method for obtaining pure recombinant human prolactin
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Protein Expression and Purification
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Prolactin is a pituitary hormone that is involved diverse physiological functions, such as lactation, reproduction,
metabolism, osmoregulation, immunoregulation, and behavior. Its level of glycosylation is low in vivo, which
favors its expression in bacterial systems. In the present work recombinant human prolactin (rec-hPRL) was
expressed from the p1813-hPRL vector in Escherichia coli strain in inclusion bodies with 530.67 mg of rec-hPRL
per liter of induced bacterial culture. The solubilization and renaturation of rec-hPRL followed by two methods
described in the literature for this protein: one with detergent and basic pH, and other urea and dialyses was
done by studying. The protocol with detergent/basic pH was not successful, whereas protocol with urea/dialyses
was obtained pure protein and this was optimized. Rec-hPRL was obtained in a soluble, pure and active form,
when the sample was 8-fold concentrated in the solubilization phase, allowing 33% recovery, 3-fold more that
the original method. The pure protein was obtained with 38.37 i. u./mg activity, which is three times greater
than that of the PRL standard from the WHO. In conclusion, this work obtained the highest production of rechPRL,
and concentrating the sample eight times in the solubilization stage was decisive for obtaining a highly
concentrated, active protein for future work.
Como referenciar
AFFONSO, REGINA; SOARES, CARLOS R.; RIBELA, MARIA T.; BARTOLINI, PAOLO. High production and optimization of the method for obtaining pure recombinant human prolactin. Protein Expression and Purification, v. 152, p. 131-136, 2018. DOI: 10.1016/j.pep.2018.07.015. Disponível em: http://repositorio.ipen.br/handle/123456789/29259. Acesso em: 30 Dec 2025.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.