High throughput production of mouse monoclonal antibodies using antigen microarrays
High throughput production of mouse monoclonal antibodies using antigen microarrays
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DOI：
10.1002/pmic.200401279
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发表时间：
2005-11-01
期刊：
影响因子：
3.4
通讯作者：
Sawyer, A
中科院分区：
文献类型：
--
作者：
De Masi, F;Chiarella, P;Sawyer, A
蛋白质组学研究的最新进展强调了对高亲和力单克隆抗体的日益增长的需求，这些抗体仍然是用冗长的低通量抗体生产技术产生的。在这里，我们提出了一个半自动化，高通量的杂交瘤产生和鉴定方法。使用多重免疫、自动融合和细胞培养以及新型抗原包被的微阵列筛选测定，在6- 10周的单批运行中产生针对不同靶点的单克隆抗体。在一个大规模的实验中，其中8只小鼠分别用10种抗原免疫，我们在初次免疫后6周内产生了针对68种靶标（85%）的单克隆抗体。
Recent advances in proteornics research underscore the increasing need for high-affinity monoclonal antibodies, which are still generated with lengthy, low-throughput antibody production techniques. Here we present a semi-automated, high-throughput method of hybridoma generation and identification. Monoclonal antibodies were raised to different targets in single batch runs of 6-10wk using multiplexed immunisations, automated fusion and cell-culture, and a novel antigen-coated microarray-screening assay. In a large-scale experiment, where eight mice were immunized with ten antigens each, we generated monoclonal antibodies against 68 of the targets (85%), within 6 wk of the primary immunization.