人抗凝血酶Ⅲ纳米膜过滤工艺优化

Optimization of human antithrombin Ⅲ nanofiltration process

目的考察Asahi Kasei（旭化成）公司纳米膜过滤人抗凝血酶Ⅲ（Human AntithrombinⅢ,ATⅢ）的回收率,优化纳米膜过滤工艺。方法选取旭化成公司3种不同规格型号的纳米膜Planova~15N,Planova~20N和Planova~Bio EX进行ATⅢ过滤回收率考察,通过留取生产工艺中不同阶段不同蛋白浓度的制品,优化过滤蛋白总量,确定纳滤米过滤工艺位置。结果蛋白浓度16 mg/m L的ATⅢ通过Planova~20N过滤获得蛋白总量最高,ATⅢ蛋白及效价回收率也较高,为最佳纳滤蛋白浓度。结论通过本研究筛选出Planova~20N,最佳纳滤蛋白浓度为16 mg/m L,经去除病毒验证其对细小病毒的去除率LRV〉4.0,最终确定纳滤工艺在人抗凝血酶Ⅲ工艺中超滤浓缩后,既符合生产工艺要求并可获得高回收率。

Objective To study the nanofiltration membrane（ Asahi Kasei） recovery of human antithrombin Ⅲ（ ATⅢ） and to optimize the filtration process. Methods Three different types and specifications of nanofiltration membranes were selected,such as Planova ~15N,Planova ~20N and Planova ~Bio EX. The recovery of AT Ⅲ by filtration was investigated by taking specimens at different stages of the manufacture process with different protein concentrations. The total protein filtration amount was optimized. The location of nanofiltration in the whole manufacture process was determined. Results AT Ⅲ with protein concentration of 16 mg / m L filtrated by Planova~20N had the highest total protein amount and better protein and activity recovery. Conclusion Planova~20N was screened as the optimum nanofilter used in AT Ⅲ manufacture process. The highest recovered protein concentration was 16 mg / m L. The LRV（ Log Reduction Values） of parvovirus removal rate was not lower than 4. 0. Ultimately,nanofiltration was operated after ultrafiltration which conforms to the requirements of AT Ⅲ manufacture process,and higher recovery was achieved.