Fully automated production of potential Malaria vaccines with Pichia pastoris in integrated processing

Abstract

Here, we have studied the setup of an integrated bioprocess for the production of artificial Malaria vaccine candidates with Pichia pastoris. Production of pharmaceutically relevant proteins such as vaccines has high demands regarding protein processing in the bioreactor and for subsequent purification. To implement this challenging protein expression process, a highly instrumented bioreactor was configured for repeated fed batch cultivations and supplemented with an at‐line monitoring of the target protein production via HPLC. The integration of a fast in situ purification of the sensitive products using an expanded bed adsorption for a sequential integrated bioprocess allows cyclic product separation. Thus, a fully automated production of artificial malaria vaccines was achieved.

Diversity Covering AMA1-MSP1 19 Fusion Proteins as Malaria Vaccines

“…With approximately 30 mg/liter, the yield was slightly lower than the yield of the AMA1 unit alone (33), and optimization of the fermentation conditions for D1M1 resulted in an increased yield (40). The yield of the D1M1D2M2D3 fusion product was very low at 0.5 mg/liter, too low to produce vaccine at a large scale, for which a greater than 100-fold increase in yield would need to be achieved.…”

Sequential/parallel production of potential Malaria vaccines – A direct way from single batch to quasi-continuous integrated production

“…This process was developed for fully automated operation, where 80% fermentation broth of a daily cultivation was first harvested into a conditioning tank, where the EBA conditions were adjusted and then the broth passed into an EBA procedure. This cell release and protein capture lead to elution fractions between 65% and 98% purity, even running not optimized cultivation and purification protocols (Martens et al, 2011). The remaining 20% of cultivation broth in the production reactor was refreshed and used for a new one day production in parallel to the purification of the previous step.…”

“…The target protein in the production bioreactor was detected at-line with a HPLC measurement technique (VWR International, Germany) explained earlier by (Martens et al, 2011). In addition this bioreactor is equipped with spectroscopic measurement devices for NIR-(Polytec, Germany), Raman-(Kaiser Optics, USA) and 2D-fluorescence-spectroscopy (Delta Optics, Denmark) (Voss et al, 2014 ).…”

Optimization of a preparative multimodal ion exchange step for purification of a potential malaria vaccine

“…The biological material that was used was produced by an integrated manufacturing process consisting of a sequential cultivation of Pichia pastoris strain KM71H phenotype Mut S and a subsequent capture via Expanded Bed Adsorption EBA Immobilized Metal chelate Affinity Chromatography (IMAC) (STREAMLINE 50 column and STREAMLINE Chelating media, GE Healthcare, Uppsala, Sweden) described by Martens et al [21].…”

“…The target protein was produced in an integrated production process described by Martens et al [21]. The aim of the work presented is to develop an intermediate purification step after an ultrafiltration/diafiltration in the preparative integrated bioprocess.…”