UMR CNRS 7292, GICC, Université François Rabelais, 37200, Tours, France.
Mol Genet Genomics. 2013 Aug;288(7-8):347-63. doi: 10.1007/s00438-013-0754-8. Epub 2013 Jun 7.
The molecular domestication of several DNA transposons that occurred during the evolution of the mammalian lineage, has led to the emergence of at least 43 genes, known as neogenes. To date, the limited availability of efficient commercial antibodies directed against most of their protein isoforms hampers investigation of their expression in vitro and in situ. Since immunization protocols using peptides or recombinant proteins have revealed that it is difficult to recover antibodies, we planned to produce antisera in mice using a new technique of nanopheres/DNA immunization, the ICANtibodies™ technology. Here, we investigate the possibilities of obtaining polyclonal antibodies for 24 proteins or protein domains using this immunization strategy. We successfully obtained 13 antisera that were able to detect neogenic proteins by Western blotting and ELISA in protein extracts of transiently-transfected cells and various cancer cell lines, plus another two that only detected the in ELISA and in in situ hybridizations. The features required for the production of these antibodies are analyzed and discussed, and examples are given of the advantages they offer for the study of neogenic proteins.
哺乳动物谱系进化过程中，几种 DNA 转座子的分子驯化导致了至少 43 个基因的出现，这些基因被称为新基因。迄今为止，由于缺乏针对其大多数蛋白质同工型的有效商业抗体，限制了它们在体外和原位的表达研究。由于使用肽或重组蛋白的免疫方案表明，回收抗体很困难，因此我们计划使用一种新的纳米颗粒/DNA 免疫技术，即 ICANtibodies™ 技术，在小鼠中产生抗血清。在这里，我们使用这种免疫策略研究了获得针对 24 种蛋白质或蛋白质结构域的多克隆抗体的可能性。我们成功获得了 13 种抗血清，这些抗血清能够通过 Western blot 和 ELISA 在瞬时转染细胞和各种癌细胞系的蛋白质提取物中检测到新基因蛋白，另外还有两种仅在 ELISA 和原位杂交中检测到。我们分析和讨论了产生这些抗体所需的特征，并给出了它们在新基因蛋白研究中的优势示例。
