Rathore Anurag S, Bilbrey Robert E, Steinmeyer David E
Bioprocess and Formulation Technology, Pharmacia Corporation, 700 Chesterfield Parkway North, Chesterfield, Missouri 63198, USA.
Biotechnol Prog. 2003 Sep-Oct;19(5):1541-6. doi: 10.1021/bp034030s.
Small-scale experiments were performed to evaluate the impact of several processing variables on recovery by osmotic shock of a protein product expressed in E. coli utilizing feed material manufactured at large scale. Equilibration time, type and concentration of osmotic supplements, and pH of the osmotic shock buffer were found to have significant impact on the osmotic shock recovery. Improved recovery was documented under conditions of increased equilibration time before the osmotic shock and increased Tris-HCl concentration in the osmotic shock buffer, as well as initial dispersal of cells into the osmotic shock buffer. Recovery of up to 65% was achieved under the optimized conditions.
进行了小规模实验，以评估几个加工变量对利用大规模生产的进料在大肠杆菌中表达的蛋白质产品通过渗透休克法回收的影响。发现平衡时间、渗透补充剂的类型和浓度以及渗透休克缓冲液的pH值对渗透休克回收有显著影响。在渗透休克前平衡时间增加、渗透休克缓冲液中Tris-HCl浓度增加以及细胞初始分散到渗透休克缓冲液的条件下，回收率得到了提高。在优化条件下实现了高达65%的回收率。
