Patent ID: 6599697
Filing Date: 2003-07-29
Classification: C07K,C12N,C40B

Abstract:
A process for producing a multi-combinatorial phagemid library comprising the steps of:a) providing a first repertoire of genes encoding a population of either type of antibody VL regions or antibody VH regions, and b) providing at least one gene encoding a variable region of the other type of VL regions or VH regions, or a second repertoire of genes encoding a population of variable regions of the other type of VL regions or VH regions; introducing the genes of the first repertoire into a first vector in order to form a population of vectors carrying different genes of the first repertoire, and introducing the gene encoding the other type of light or heavy antibody chain or the genes of the second repertoire into a second vector, wherein at least one of the vectors, termed recipient, is a phagemid and the vectors respectively contain the E. coli AttB and the phage &lgr; AttP specific recombination sites which are arranged in such a manner as to permit recombination under the influence of the associated recombinase or integrase; and recombining said vectors under the influence of said recombinase or integrase, thereby forming, in a single recombinant vector resulting from the recombination, stable junction sequences and enabling said recombinant vector to contain, in an irreversible manner, a gene of a variable region of either the light or heavy chain antibody types and a gene of the variable region of the other type of light or heavy antibody chain, and to express the two genes in the form of linked polypeptides which are able to appear on the external surface of the product of the said vector, being maintained there and being linked together in a multimeric manner, or simulating a multimer; wherein the recombinant vector is arranged to have a selection marker which is initially non-functional and which is rendered functional by the recombination.