Moutel Sandrine, El Marjou Ahmed, Vielemeyer Ole, Nizak Clément, Benaroch Philippe, Dübel Stefan, Perez Franck
CNRS UMR144, 26 rue d'Ulm, F75248 Paris Cedex 05, France.
BMC Biotechnol. 2009 Feb 26;9:14. doi: 10.1186/1472-6750-9-14.
Genomic, transcriptomic and proteomic projects often suffer from a lack of functional validation creating a strong demand for specific and versatile antibodies. Antibody phage display represents an attractive approach to select rapidly in vitro the equivalent of monoclonal antibodies, like single chain Fv antibodies, in an inexpensive and animal free way. However, so far, recombinant antibodies have not managed to impose themselves as efficient alternatives to natural antibodies.
We developed a series of vectors that allow one to easily fuse single chain Fv antibodies to Fc domains of immunoglobulins, improving their sensitivity and facilitating their use. This series enables the fusion of single chain Fv antibodies with human, mouse or rabbit Fc so that a given antibody is no longer restricted to a particular species. This opens up unlimited multiplexing possibilities and gives additional value to recombinant antibodies. We also show that this multi-Fc species production system can be applied to natural monoclonal antibodies cloned as single chain Fv antibodies and we converted the widely used 9E10 mouse anti-Myc-tag antibody into a human and a rabbit antibody.
Altogether, this new expression system, that brings constant quality, sensitivity and unique versatility, will be important to broaden the use of recombinant and natural monoclonal antibodies both for laboratory and diagnosis use.
基因组、转录组和蛋白质组学项目常常缺乏功能验证,这使得对特异性和多功能抗体的需求极为迫切。抗体噬菌体展示是一种颇具吸引力的方法,能够以低成本且无需动物实验的方式,在体外快速筛选出相当于单克隆抗体的物质,如单链Fv抗体。然而,迄今为止,重组抗体尚未成功成为天然抗体的有效替代物。
我们开发了一系列载体,可轻松将单链Fv抗体与免疫球蛋白的Fc结构域融合,提高其敏感性并便于使用。该系列载体能够使单链Fv抗体与人类、小鼠或兔的Fc结构域融合,从而使特定抗体不再局限于某一特定物种。这开启了无限的多重检测可能性,并赋予重组抗体额外价值。我们还表明,这种多Fc物种生产系统可应用于克隆为单链Fv抗体的天然单克隆抗体,并且我们将广泛使用的9E10小鼠抗Myc标签抗体转化为了人源和兔源抗体。
总之,这种新的表达系统具有恒定的质量、敏感性和独特的多功能性,对于扩大重组和天然单克隆抗体在实验室和诊断应用中的使用将具有重要意义。