Purification of therapeutic proteins mediated by in vivo polyester immobilized sortase.
Biotechnol Lett
; 40(2): 369-373, 2018 Feb.
Article
em En
| MEDLINE
| ID: mdl-29124516
ABSTRACT
OBJECTIVES:
To overcome laborious and costly procedures often associated with therapeutic protein production and purification, in vivo polyester immobilized sortase is explored for the production of human tumor necrosis factor alpha (TNFα) and human interferon alpha 2b (IFNα2b) by Escherichia coli.RESULTS:
Hybrid genes encoding PhaC-Sortase-TNFα or PhaC-Sortase-IFNα2b fusions (with a LPETG recognition signal immediately before TNFα or IFNα2b), mediated intracellular production of polyester (polyhydroxyalkanoate, PHA) beads in Escherichia coli. Upon isolation of respective PHA beads, pure soluble TNFα or IFNα2b was released by activating sortase via addition of CaCl2 and triglycine. TNFα and IFNα2b each were recognized by corresponding conformational antibodies in an ELISA assay.CONCLUSIONS:
In vivo polyester immobilized sortase could be exploited for production and purification of high-value therapeutic proteins without laborious and costly downstream processing.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Poliésteres
/
Proteínas de Bactérias
/
Proteínas Recombinantes de Fusão
/
Cisteína Endopeptidases
/
Aminoaciltransferases
/
Enzimas Imobilizadas
Limite:
Humans
Idioma:
En
Revista:
Biotechnol Lett
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Nova Zelândia