Patent ID: 8101553
Filing Date: 2012-01-24
Classification: C07K,C12N

Abstract:
1. A method for preparing a gene library comprising combinations of different light chain variable region genes and heavy chain variable region genes of immunoglobulin, the method comprising: (a) selecting a plurality of human derived different light chain variable region genes encoding light chain molecules, wherein the light chain variable region genes are selected by detecting a holding structure of a complex through an association of a heavy chain variable chain molecule with a light chain variable chain molecule to form Fab, further wherein the association is detected by both an antibody of I) and an antibody of II) binding to Fab, when the light chain molecule is combined with an expression product of a heavy chain variable region gene encoding the heavy chain variable chain molecule, wherein I) is an antibody against the light chain, and II) is an antibody against the heavy chain or Fab, and wherein the plurality of different light chain variable region genes is 100 for each of κ chain genes and λ chain genes; (b) recovering each of the human derived different light chain variable region gene selected in step (a) from the heavy chain variable region gene; (c) constructing a gene library consisting of a collection of the human derived different light chain variable region genes recovered in step b); and (d) combining the gene library of light chain variable region genes obtained in step (c) with one or more libraries of genes encoding human derived heavy chain variable regions having a full repertoire of heavy chain variable region genes, whereby the resulting gene library maintains diversity in the heavy chain variable region.