title: Bioprocess considerations for expanded-bed chromatography of crude canola extract: sample preparation and adsorbent reuse.
abstract: Compared to the conventional microbial and mammalian systems, transgenic plants produce proteins in a different matrix. This provides opportunities and challenges for downstream processing. In the context of the plant host Brassica napus (canola), this work addresses the bioprocessing challenges of solid fractionation, resin fouling by native plant components (e.g., oil, phenolics, etc.), hydrodynamic stability, and resin reuse for expanded bed adsorption for product capture. Plant tissue processing and subsequent protein extraction typically result in an extract with a high content of solids containing a wide particle-size distribution. Without removal of larger particles, the column inlet distributor plugged. The larger particles (> 50 microm) were easily removed through centrifugal settling comparable to that attainable with a scroll decanter. The remaining solids did not affect the column performance. Less than 4% of the lipids and phenolics in the fed extract bound to STREAMLINE trade mark DEAE resin, and this small proportion could be satisfactorily removed using recommended clean-in-place (CIP) procedures. Hydrodynamic expansion and adsorption kinetics of the STREAMLINE trade mark DEAE resin were maintained throughout 10 cycles of reuse, as was the structural integrity of the resin beads. No significant accumulation of N-rich (e.g., proteins) and C/O-rich components (e.g., oil and phenolics) occurred over the same period.
authors.affiliation: Chemical Engineering Department, 2114 Sweeney Hall, Iowa State University, Ames, Iowa 50011, USA.
authors.forename: Yun
authors.initials: Y
authors.lastname: Bai
authors.forename: Charles E
authors.initials: CE
authors.lastname: Glatz
mesh_terms: D029688:Brassica napus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002852:Chromatography, Ion Exchange / Q000379:methods*; D019544:Equipment Failure Analysis; D017743:Equipment Reuse; D004983:Ethanolamines; D010316:Particle Size; D010936:Plant Extracts / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification*; D012639:Seeds / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014462:Ultrafiltration / Q000379:methods
chemical_list: D004983:Ethanolamines; D010936:Plant Extracts; D011994:Recombinant Proteins; C007369:2-diethylaminoethanol