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Separation Science and Technology, Vol.48, No.13, 1913-1922, 2013
Pilot-Scale Recovery of Phycoerythrin from Porphyridium cruentum using Expanded Bed Adsorption Chromatography
In this paper, a process was developed on a pilot scale for the recovery of B-phycoerythrin from Porphyridium cruentum using EBA. The process was optimized on a small scale using a 15mm ID column, then scaled up to 150mm ID columns. In the developed process, the phycobiliproteins were extracted by osmotic shock and then separated by applying the centrifuged cell suspension to an EBA column. Following this, the PE-rich solution was eluted to final purity using packed bed chromatography. Optimal conditions to separate B-phycoerithrin using EBA on a small scale were: 0.88mg B-PE/mL adsorbent sample load, H/H-0=2 and a sample viscosity of 1.068 mP. The EBA process was then scaled up by increasing the ID column (15, 25, 40, 60, and 90mm) and finally the process was developed on a pilot scale using a 150mm ID column (a scale-up factor of 100). The yield from the EBA step ranged between 71-78%, whichever ID column was used. The overall purification process yield was 54%, purification steps were monitored using SDS-PAGE, whereas protein purity was confirmed using spectroscopy. Results show that EBA is a scalable technology that allows large quantities of B-PE to be obtained, thus reducing product loss and maintaining high protein recovery while reducing both processing cost and time.
Keywords:anion-exchange chromatography;B-phycoerythrin;expanded bed adsorption chromatography;Porphyridium cruentum;phycobiliproteins;preparative protein purification