Production of human alpha-1-antitrypsin from transgenic rice cell culture in a membrane bioreactor

Biotechnol Prog. 2005 May-Jun;21(3):728-34. doi: 10.1021/bp0496676.

Abstract

Transgenic plant cell cultures offer a number of advantages over alternative host expression systems, but so far relatively low product concentrations have been achieved. In this study, transgenic rice cells are used in a two-compartment membrane bioreactor (CELLine 350, Integra Biosciences) for the production of recombinant alpha-1-antitrypsin (rAAT). Expression of rAAT is controlled by the rice alpha-amylase (RAmy3D) promoter, which is induced in the absence of sugar. The extracellular product is retained in the bioreactor's relatively small cell compartment, thereby increasing product concentration. Due to the packed nature of the cell aggregates in the cell compartment, a clarified product solution can be withdrawn from the bioreactor. Active rAAT reached levels of 100-247 mg/L (4-10% of the total extracellular protein) in the cell compartment at 5-6 days postinduction, and multiple inductions of the RAmy3D promoter were demonstrated.

Publication types

  • Evaluation Study
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bioreactors*
  • Cell Culture Techniques / instrumentation*
  • Cell Culture Techniques / methods
  • Cell Line
  • Equipment Design
  • Equipment Failure Analysis
  • Humans
  • Membranes, Artificial*
  • Oryza / physiology*
  • Plants, Genetically Modified / growth & development
  • Plants, Genetically Modified / metabolism
  • Protein Engineering / instrumentation
  • Protein Engineering / methods*
  • Recombinant Proteins / biosynthesis
  • alpha 1-Antitrypsin / biosynthesis*
  • alpha 1-Antitrypsin / genetics

Substances

  • Membranes, Artificial
  • Recombinant Proteins
  • SERPINA1 protein, human
  • alpha 1-Antitrypsin