Analysis of some operating parameters of novel adsorbents for recovery of proteins in expanded beds.
We have analysed some operational parameters for two novel adsorbents intended for recovery of proteins from particle-containing feedstocks using expanded bed adsorption. The adsorbents tested were STREAMLINE DEAE and STREAMLINE SP, ion exchangers based on an agarose/crystalline quartz composite. Parameters analysed included bed expansion, adsorption efficiency, washing and elution. Bed expansion was considerably lower for STREAMLINE adsorbents compared to conventional agarose based media, higher flow velocities were thus possible during the expanded bed process. Breakthrough capacity was 63 mg ml-1 for lysozyme on STREAMLINE SP and 36 mg ml-1 for bovine serum albumin on STREAMLINE DEAE at a flow velocity of 300 cm h-1. To achieve high breakthrough capacity, the sedimented bed height should be at least 10 cm. Furthermore, breakthrough capacity increased to some extent when temperature was increased from room temperature to 36 degrees C, a phenomenon which can be useful in some processes. The number of living E. coli cells in the effluent was reduced by a factor of 10(5) after washing with 15 sedimented bed volumes. The optimal flow velocity for elution was 100 cm h-1 considering time for elution and volume of the eluted fraction. Flow direction during elution in packed bed mode had little impact on the elution volume, however, elution in expanded bed mode increased the volume by approx. 40%. The data presented on the performance of STREAMLINE adsorbents show that they are very useful for recovery of proteins from particle-containing feedstocks using expanded bed adsorption.
Pharmacia Biotech AB, Uppsala, Sweden.
R
R
Hjorth
S
S
Kämpe
M
M
Carlsson
D000327:Adsorption; D000818:Animals; D001672:Biocompatible Materials; D002417:Cattle; D002477:Cells; D002850:Chromatography, Gel / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004926:Escherichia coli / Q000302:isolation & purification; D009113:Muramidase / Q000302:isolation & purification; D010316:Particle Size; D011506:Proteins / Q000302:isolation & purification*; D011791:Quartz; D011994:Recombinant Proteins / Q000302:isolation & purification; D012685:Sepharose; D012710:Serum Albumin, Bovine / Q000302:isolation & purification; D013696:Temperature
D001672:Biocompatible Materials; D011506:Proteins; D011994:Recombinant Proteins; D011791:Quartz; C097574:sepharose fast flow SP; D012710:Serum Albumin, Bovine; D012685:Sepharose; D009113:Muramidase