Trinh Loc, Phue Je-Nie, Jaluria Pratik, Tsai Chiawei W, Narum David L, Shiloach Joseph
Biotechnology Unit, NIDDK, NIH Bldg 14A, Room 173, Bethesda, MD 20892, USA.
Biotechnol Lett. 2006 Jul;28(13):951-8. doi: 10.1007/s10529-006-9029-3. Epub 2006 Jun 20.
An experimental malaria transmission blocking vaccine antigen, Pfs25H, expressed and secreted from Pichia pastoris was recovered and purified using a screenless expanded bed column equipped with a rotating fluid distribution system. This column was able to accommodate feed stock, containing 30% biomass, at a flow rate of 300-400 cm/h without affecting column stability. This capability is three times higher than the capability of the expanded bed column currently in use, which is equipped with a perforated plate fluid distribution system; this design could accommodate biomass concentrations of only up to 10%. The screen-less design did not affect the binding capacity, purification level or process yield and, therefore, shorten the process. Purified Pfs25H of 6.4 g were recovered from 37 l of Pichia pastoris culture in one step.
一种从毕赤酵母中表达并分泌的实验性疟疾传播阻断疫苗抗原Pfs25H，使用配备旋转流体分配系统的无筛膨胀床柱进行回收和纯化。该柱能够以300 - 400厘米/小时的流速容纳含有30%生物质的进料，而不影响柱的稳定性。这种能力比目前使用的配备多孔板流体分配系统的膨胀床柱高两倍；这种设计只能容纳高达10%的生物质浓度。无筛设计不影响结合能力、纯化水平或工艺产率，因此缩短了工艺。一步从37升毕赤酵母培养物中回收了6.4克纯化的Pfs25H。
