Université Paris Descartes; Paris, France; ENSCP Chimie ParisTech; Paris, France; CNRS UMR8151; Paris, France; Inserm U1022; Paris, France.
Unité des Bactéries Anaérobies et Toxines; Paris, France.
Hum Vaccin Immunother. 2013 Oct;9(10):2147-56. doi: 10.4161/hv.25192. Epub 2013 Jul 22.
Raising high titer antibodies in animals is usually performed by protein immunization, which requires the long and sometimes difficult step of production of the recombinant protein. DNA immunization is an alternative to recombinant proteins, only requiring the building of an eukaryotic expression plasmid. Thanks to efficient DNA delivery techniques such as in vivo electroporation, DNA vaccination has proven useful the last few years. In this work, we have shown that it is possible to raise very high antibody titers in rabbit by DNA electroporation of an antigen encoding plasmid in the skeletal muscle with the right set of electrodes and rabbit strain. In a model of botulinum toxins types A and E, the neutralizing titers obtained after three treatments were high enough to fit the European Pharmacopeia, while it did not for type B toxin. Furthermore, the raised antibodies have high avidity and are suitable for in vitro and in vivo immunodetection of proteins.
在动物体内产生高滴度的抗体通常通过蛋白质免疫来实现,这需要进行重组蛋白的长时间且有时困难的生产步骤。DNA 免疫是重组蛋白的一种替代方法,只需要构建真核表达质粒。由于电穿孔等有效的 DNA 递送技术,DNA 疫苗在过去几年中已被证明非常有用。在这项工作中,我们已经表明,通过在骨骼肌中用电穿孔的方法将编码抗原的质粒 DNA 转染到合适的一组电极和兔子品系中,有可能在兔子体内产生非常高的抗体滴度。在 A 型和 E 型肉毒毒素的模型中,经过三次处理后获得的中和滴度足以符合欧洲药典的要求,而 B 型毒素则不行。此外,所产生的抗体具有高亲和力,适用于蛋白质的体外和体内免疫检测。
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