Antibody arrays for high-throughput screening of antibody-antigen interactions
Antibody arrays for high-throughput screening of antibody-antigen interactions
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DOI：
10.1038/79494
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发表时间：
2000-09-01
影响因子：
46.9
通讯作者：
Tomlinson, IM
中科院分区：
文献类型：
--
作者：
de Wildt, RMT;Mundy, CR;Tomlinson, IM
我们已经开发了一种基于创建抗体阵列的高通量筛选重组抗体的新技术。我们的方法采用机器人筛选和高密度网格法对含有抗体基因的细菌进行筛选，然后进行基于过滤器的酶联免疫吸附试验（ELISA）筛选，以鉴定表达结合抗体片段的克隆。通过消除液体处理的需要，我们可以一次筛选多达18342种不同的抗体克隆，并且由于克隆来自主库，相同的抗体可以被双重发现，同时针对15种不同的抗原进行筛选。我们已经在几个不同的应用中使用了我们的技术，包括分离针对不纯蛋白质和复杂抗原的抗体，其中几轮噬菌体展示经常失败。我们的研究结果表明，抗体阵列可以用于鉴定差异表达蛋白。
We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins.