A microengraving method for rapid selection of single cells producing antigen-specific antibodies
A microengraving method for rapid selection of single cells producing antigen-specific antibodies
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DOI：
10.1038/nbt1210
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发表时间：
2006-06-01
影响因子：
46.9
通讯作者：
Ploegh, Hidde L.
中科院分区：
文献类型：
--
作者：
Love, J. Christopher;Ronan, Jehnna L.;Ploegh, Hidde L.
识别感兴趣的特定抗原的单克隆抗体被用作治疗剂和生物医学研究的工具(1)。发现单个单克隆抗体需要从产生多种特异性抗体的细胞的多克隆混合物中检索单个杂交瘤。然而，通过有限的连续稀释分离杂交瘤所需的时间限制了可用抗体的多样性和广度。在这里，我们提出了一种基于凹版印刷的软光刻方法，以产生包含单细胞分泌产物的微阵列。这些雕刻阵列能够快速（< 12小时）和高通量（100,000个单个细胞）系统，用于鉴定，恢复和克隆扩增产生抗原特异性抗体的细胞。原则上，该方法可适用于以多路复用方式检测任何分泌产物。
Monoclonal antibodies that recognize specific antigens of interest are used as therapeutic agents and as tools for biomedical research(1). Discovering a single monoclonal antibody requires retrieval of an individual hybridoma from polyclonal mixtures of cells producing antibodies with a variety of specificities. The time required to isolate hybridomas by a limiting serial-dilution, however, has restricted the diversity and breadth of available antibodies. Here we present a soft lithographic method based on intaglio printing to generate microarrays comprising the secreted products of single cells. These engraved arrays enable a rapid (< 12 h) and high-throughput (> 100,000 individual cells) system for identification, recovery and clonal expansion of cells producing antigen-specific antibodies. This method can be adapted, in principle, to detect any secreted product in a multiplexed manner.