Pilot Scale Recovery of Phycocyanin from Spirulina platensis Using Expanded Bed Adsorption Chromatography

The use of expanded bed adsorption chromatography (EBAC) for pilot scale recovery of C-phycocyanin (C-PC) from the cyanobacteria Spirulina platensis is described. The description includes the whole procedure, from small-scale method optimization to pilot scale. Initially, the phycobiliproteins are extracted by osmotic shock and separated by applying the centrifuged cell suspension to an EBAC column, using Streamline-DEAE as adsorbent. Next, the PCs rich solution is eluted to finally be purified by packed bed chromatography using DEAE-cellulose. The EBAC operation was optimized on a small scale using a 15 mm internal diameter (ID) column. The optimal conditions were: a sample load of 0.91 mg C-PC mL(-1) adsorbent, H/H (0) = 2 and a sample viscosity of 1.084 mP. The EBAC process was then scaled up by increasing the column ID (15, 25, 40, 60 and 90 mm) and, finally, the process was developed at pilot scale using a 150 mm ID column obtaining a scale-up factor of 100. The yield of the EBAC step is in the range of 75-79% whatever the column diameter. To obtain pure C-PC, conventional chromatography with DEAE-cellulose was utilized. The overall yield of the purification process after all steps is 59%. The purification steps are monitored using SDS-PAGE and the purity of recovered C-PC is confirmed by absorption and emission spectroscopy. Results show that EBAC method is a scalable technology that allows large quantities of C-PC to be obtained without product loss, maintaining a high protein recovery while reducing both processing cost and times.