Hyseq Pharmaceuticals, 675 Almanor Ave., Sunnyvale, CA, 94086, USA.
Cytotechnology. 2002 Jan;38(1-3):43-6. doi: 10.1023/A:1021141712344.
UCOE vectors contain non-tissue specific chromatin-opening-elements that permit rapid expression of a protein in anintegration independent manner. Efficient expression can bederived from a single copy of an integrated gene site resulting ina higher percentage of cells expressing the marker gene in theselected pool in comparison to standard non-UCOE containingvectors. This, in combination with the utilization of a serum-free, suspension adapted parent cell line allows for rapidproduction of large quantities of protein in a short period oftime. Utilizing this system more than 300 mg of a recombinantantibody has been produced in less than 1 month from transfectionpools in shake flask. Selected subclones have been scaled intosmall bioreactors in less than 2 months, producing significantquantities of monoclonal antibody using a protocol generic for theparent cell line. The increased efficiency obtained with the UCOEvector reduces the number of transfectants which need to bescreened in order to obtain high productivity subclones.Transfection of a standard host cell line, preadapted to grow in alarge-scale setting, allows for rapid cell line developmentdecreasing the transition time from research into development andmanufacturing. Alternatively, the traditional approach of using aparent cell line which requires serum-free and suspensionadaptation after transfection further increases the need forscreening a large number of subclones, because many of thesubclones will not be able to grow under conditions that allowlarge-scale protein production. The use of a preadapted cell linecan reduce the time required to develop a cell line from months toweeks.
UCOE 载体包含非组织特异性染色质开放元件,可使蛋白质在独立于整合的方式下快速表达。高效表达可源于整合基因位点的单个拷贝,与包含标准非 UCOE 的载体相比,在所选池中的表达标记基因的细胞比例更高。这与利用无血清、悬浮适应的亲本细胞系相结合,允许在短时间内快速生产大量蛋白质。利用该系统,在摇瓶转染池内不到 1 个月就生产出了超过 300 毫克的重组抗体。选定的亚克隆已在不到 2 个月的时间内扩大到小型生物反应器中,使用适用于亲本细胞系的通用方案生产出了大量单克隆抗体。UCOE 载体获得的效率提高减少了需要筛选的转染体数量,以获得高生产力的亚克隆。对预先适应大规模生长的标准宿主细胞系进行转染,可快速开发细胞系,减少从研究到开发和制造的过渡时间。或者,使用需要转染后无血清和悬浮适应的亲本细胞系的传统方法进一步增加了筛选大量亚克隆的需求,因为许多亚克隆将无法在允许大规模蛋白质生产的条件下生长。使用预先适应的细胞系可以将细胞系的开发时间从数月缩短至数周。