Patent ID: 8623594
Filing Date: 2014-01-07
Classification: C07K,C12N,C40B

Abstract:
1. A method of isolating a DNA encoding a protein which binds to a platelet antigen, said method comprising: generating a shuffled library comprising: 1) mixing DNAs from at least a first γκ phage display library and a second γλ phage display library, wherein said first γκ phage display library comprises at least one virus vector that comprises DNA encoding a heavy and light chain isolated from panning a population of phage against a platelet antigen and said second γλ phage display library comprises at least one virus vector that comprises DNA encoding a heavy and light chain isolated from panning a population of phage against said platelet antigen, further wherein said panning a population of phage against said platelet antigen comprises adding a magnetic label to cells bearing said platelet antigen on their surface, thereby producing magnetically labeled cells; incubating said phage display library with said magnetically labeled cells in the presence of an excess of unlabeled cells which do not express said platelet antigen to form a mixture, whereby said at least one virus vector binds to said magnetically labeled cells; isolating magnetically labeled cells from said mixture, whereby said at least one virus vector is isolated from said mixture, and obtaining DNA encoding a heavy and light chain from said isolated virus vector; 2) dissociating said DNA encoding the heavy chains from said at least one virus vector from said first γκ phage display library and said at least one virus vector from said second γλ phage display library using a restriction endonuclease thereby generating a mixture of dissociated heavy chain DNA segments and dissociated vectors encoding a light chain and re-ligating the dissociated heavy chain DNA segment into a dissociated vector encoding a light chain thereby generating a hybrid virus vector encoding a light chain and a heavy chain that is not encoded by a single vector in the first and second phage display library; incubating said hybrid virus vector with magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said platelet antigen to form a mixture, whereby said hybrid virus vector binds to said magnetically labeled cells; isolating magnetically labeled cells from said mixture, whereby said hybrid virus vector is isolated from said mixture; and isolating said DNA encoding said protein from said isolated hybrid virus vector.