Integrated approach to produce a recombinant, his-tagged human α-galactosidase a in mammalian cells

José Luis Corchero, Rosa Mendoza, Julia Lorenzo, Victor Rodríguez-Sureda, Carmen Domínguez, Esther Vázquez, Neus Ferrer-Miralles, Antonio Villaverde

Research output: Contribution to journalArticleResearchpeer-review

9 Citations (Scopus)

Abstract

Successful production of recombinant proteins (r-proteins) by transient gene expression (TGE) depends on several parameters (including producer cells, culture conditions, transfection procedure, or expression vector) that should be optimized when producing any recombinant product. In this work, TGE-based production of human α-galactosidase A (GLA) is described. Producer cells, expression vectors, and parameters influencing cell metabolism after transfection have been tested. The enzyme is secreted, has the right molecular weight, and is enzymatically active. Productivities of up to 30-40 mg/L have been achieved, with a simple, fast procedure. A 6 × His tag allows enzyme purification in a single step, rendering a highly pure product. We propose a TGE-based protocol able to produce up to several milligrams per liter of highly pure, active GLA in a time as short as a few days. By this, enough amounts of engineered versions of the enzyme can be easily produced to be tested in vitro or in preclinical trials. © 2011 American Institute of Chemical Engineers (AIChE).
Original languageEnglish
Pages (from-to)1206-1217
JournalBiotechnology Progress
Volume27
DOIs
Publication statusPublished - 1 Oct 2011

Keywords

  • Fabry's disease
  • Human α-galactosidase A
  • Mammalian cells
  • Recombinant protein
  • Transient gene expression

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