Brobjer M
Pharmacia & Upjohn Process R&D, Stockholm, Sweden.
Bioseparation. 1999;8(1-5):219-28.
A capture step was developed using the expanded bed adsorption technology to separate a protein of interest on a cation exchanger from a crude Escherichia coli homogenate. This method was developed in bench-top scale using a STREAMLINE 25 column (Amersham Pharmacia Biotech, Sweden) and STREAMLINE SP. The development was based on earlier experiments performed in a packed bed column (SP-Sepharose FF) to investigate the conditions for sample application, wash and elution. The packed bed method was transformed into an expanded bed method by slightly modifying the wash procedure and cleaning in place (CIP). This method was then scaled-up to pilot scale and used for production of the fusion protein according to cGMP. The yield over the step in pilot scale was 70-85% compared with only 30-50% in small scale. Pressure build-up, attachment of biomass to the adsorbent and collapses of the expanded bed were phenomena seen in small scale but not in pilot scale. The scale-up of the step significantly improved the performance of the step.
采用扩张床吸附技术开发了一种捕获步骤，用于从大肠杆菌粗匀浆中在阳离子交换剂上分离目标蛋白质。该方法在台式规模上使用STREAMLINE 25柱（瑞典Amersham Pharmacia Biotech公司）和STREAMLINE SP进行开发。该开发基于早期在填充床柱（SP-Sepharose FF）中进行的实验，以研究样品上样、洗涤和洗脱的条件。通过略微修改洗涤程序和在位清洗（CIP），将填充床方法转变为扩张床方法。然后将该方法放大到中试规模，并根据cGMP用于融合蛋白的生产。中试规模下该步骤的产率为70-85%，而小规模下仅为30-50%。小规模时出现了压力升高、生物质附着在吸附剂上以及扩张床塌陷等现象，而中试规模时未出现。该步骤的放大显著提高了该步骤的性能。
