Aldrich Teri L, Viaje Aurora, Morris Arvia E
Department of Cell Sciences, Amgen, Inc., 51 University Street, Seattle, Washington 98101, USA.
Biotechnol Prog. 2003 Sep-Oct;19(5):1433-8. doi: 10.1021/bp034056j.
Over the past 10 years, monoclonal antibodies and antibody fragments have become an increasingly important source of therapeutic molecules in the biotechnology industry. Drug development strategies rely on screening large numbers of candidate molecules in search of an optimized drug candidate. This strategy requires efficient production of ten to a few hundred milligrams of candidate molecules for screening in bioassays and animal models. Typically, this amount of recombinant protein expression involves large numbers of transient transfections or cloning of a recombinant cell line. Both of these approaches are time-consuming and labor-intensive. In this report, we describe the application of an EASE vector system that is capable of generating stable pools of transfected Chinese hamster ovary cells. These pooled populations of cells produce high quantities of antibody candidates without labor-intensive cloning in a 3-5 week time frame. When an optimal drug candidate has been selected, pools generated with EASE-containing vectors can also be used in subsequent cloning steps to make cell lines with improved expression levels. We demonstrate that EASE increases expression in nonamplified pools in addition to increasing amplification and viability of clonal cell lines generated with the EASE-containing vectors compared with pools and cell lines generated without EASE.
在过去10年中，单克隆抗体和抗体片段已成为生物技术产业中治疗性分子的一个越来越重要的来源。药物开发策略依赖于筛选大量候选分子以寻找优化的候选药物。该策略需要高效生产10至几百毫克的候选分子，用于生物测定和动物模型中的筛选。通常，这种数量的重组蛋白表达涉及大量的瞬时转染或重组细胞系的克隆。这两种方法都既耗时又费力。在本报告中，我们描述了一种EASE载体系统的应用，该系统能够产生稳定的转染中国仓鼠卵巢细胞库。这些细胞库群体在3至5周的时间内无需费力的克隆即可产生大量的候选抗体。当选择了最佳候选药物时，用含EASE载体产生的细胞库也可用于后续的克隆步骤，以制备具有更高表达水平的细胞系。我们证明，与不含EASE产生的细胞库和细胞系相比，EASE除了增加用含EASE载体产生的克隆细胞系的扩增和活力外，还能增加未扩增细胞库中的表达。
