Download Cloning and Expression in Pichia pastoris (Gene to Product)

Cloning and Expression in Pichia pastoris
(Gene to Product)

CBB offers following services cloning into pPIC series of commercially
available expression vectors. These vectors use methanol-responsive
AOX1 promoter and transcription terminator and secretory signal

Gene of interest is integrated into Pichia genome with PCR analysis to
confirm expected integration.

His+ or antibiotic (e.g. zeocin) based selection of transformants

Second level selection for multiple copy insertion of gene of interest for
higher expression and secretion

Expression analysis of clones from secondary screening for protein
expression and quantitation followed by selection of best expressing
clones at the shake flask level.

CBB offers both non-GMP and cGMP compliant master and working cell
banks of the production clone.

With more than 20 years fermentation experience CBB offers Pichia
pastoris fermenations up to 1000-L. Recently completed three methanol
fed Pichia runs at 500-L scale.

Since most of the proteins in Pichia are designed to secrete, CBB offers
comprehensive downstream services which includes semi-continuous
(tubular) and continuous centrifugation (Disc Stack), TFF by hollow fiber or
cassette systems. Additional purification by various chromatography
methods (Affinity, IEX, SEC and Mixed mode) are also performed for high
purity.