title: Production of recombinant protein in Pichia pastoris by fermentation.
abstract: This protocol is applicable to recombinant protein expression by small-scale fermentation using the Pichia pastoris expression system. P. pastoris has the capacity to produce large quantities of protein with eukaryotic processing. Expression is controlled by a methanol-inducible promoter, which allows a biomass-generation phase before protein production is initiated. The target protein is secreted directly into a protein-free mineral salt medium, and is relatively easy to purify. The protocol is readily interfaced with expanded bed adsorption for immediate capture and purification of recombinant protein. The setting up of the bioreactor plus the fermentation itself takes 1 wk. Making the master and user seed lots takes approximately 2 wk for each individual clone.
authors.affiliation: Department of Oncology, Royal Free and University College Medical School, University College London, Rowland Hill Street, London NW3 2PF, UK. b.tolner@ucl.ac.uk
authors.forename: Berend
authors.initials: B
authors.lastname: Tolner
authors.forename: Lisa
authors.initials: L
authors.lastname: Smith
authors.forename: Richard H J
authors.initials: RH
authors.lastname: Begent
authors.forename: Kerry A
authors.initials: KA
authors.lastname: Chester
mesh_terms: D019149:Bioreactors; D005285:Fermentation / Q000502:physiology*; D015870:Gene Expression; D010843:Pichia / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins