公开(公告)号：US20050059053A1

公开(公告)日：2005-03-17

申请号：US10895399

申请日：2004-07-21

申请人： Rainer Fischer ,  Markus Sack ,  Carmen Vaquero-Martin

发明人： Rainer Fischer ,  Markus Sack ,  Carmen Vaquero-Martin

IPC分类号： C07H21/04 ,  C07K14/705 ,  C07K16/30 ,  C12N15/62 ,  C12N15/82 ,  C12P21/02 ,  C12Q1/68 ,  C12P21/04 ,  G01N33/53

CPC分类号： C12N15/62 ,  C07H21/04 ,  C07K14/70503 ,  C07K16/3007 ,  C07K2317/13 ,  C07K2317/622 ,  C07K2317/626 ,  C07K2319/033 ,  C07K2319/21 ,  C07K2319/40 ,  C12N15/8257 ,  C12N15/8258 ,  C12P21/02

摘要： A method is described for altering the properties such as the accumulation, the stability and/or integrity, the subcellular localisation, the post-translational modifications, the ability to get purified, and the phase partitioning behaviour of natural or recombinant target proteins expressed in a host organism. The method involves the co-expression of natural or recombinant proteins along with a specific binding partner that sequesters the target recombinant protein into a complex. The binding partner is supplied as a separate protein allowing formation of intermolecular complexes or is fused to the protein of interest, allowing the formation of intramolecular complexes. The binding partner can also be used to alter the subcellular localisation without modifying the sequence or structure of the target protein itself. This can be achieved by either incorporating appropriate targeting signals into the binding ligand, which are then linked to the target protein through complex formation, or complex formation itself may alter the subcellular localisation. The same strategy can be used to provide an affinity tag to facilitate protein purification. The principle of the invention is demonstrated by the coexpression of an unstable antibody and its cognate antigen.

摘要翻译： 描述了一种用于改变诸如累积，稳定性和/或完整性，亚细胞定位，翻译后修饰，获得纯化的能力以及表达于天然或重组靶蛋白的相分离行为的性质的方法 宿主生物。 该方法包括将天然或重组蛋白与特异性结合配偶体共表达，所述特异性结合配偶体将靶重组蛋白质螯合到复合物中。 结合配偶体作为单独的蛋白质提供，允许形成分子间复合物或者与感兴趣的蛋白质融合，从而形成分子内复合物。 结合配偶体也可用于改变亚细胞定位，而不改变靶蛋白本身的序列或结构。 这可以通过将适当的靶向信号并入结合配体中来实现，然后通过复合物形成与靶蛋白连接，或者复合物形成本身可能改变亚细胞定位。 可以使用相同的策略来提供亲和标签以促进蛋白质纯化。 本发明的原理通过不稳定抗体及其同源抗原的共表达来证明。