Patent ID: 7338659
Filing Date: 2008-03-04
Classification: A61K,A61P,C07K

Abstract:
1. A method for producing an immunoglobulin containing 3 heavy chain and 3 light chain complementary determining regions from the mouse RFB4 immunoglobulin and framework sequences from human immunoglobulins comprising the steps of: a. dividing the framework sequences from the RFB4 immunoglobulin into compartmentalized sub-regions of FR1, FR2, FR3, and FR4 according to the classification of the Kabat Database; b. comparing the individual sub-region framework sequences to corresponding sequences in a collection of human immunoglobulin chains; c. selecting, from the collection, the appropriate human framework sequences to replace the original framework sequences of the FR1, FR2, FR3 and FR4 sub-regions of the RFB4 immunoglobulin, wherein the resulting immunoglobulin comprises a heavy chain variable region having the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 1 and a light chain variable region having the amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 9; d. assembling the framework sequences selected in step c; e. subcloning the assembled framework sequences of step d into heavy and light chain an expression vectors containing appropriate immunoglobulin constant heavy and light chain nucleotide sequences; f. co-transfecting mouse Sp2/0 cells with the expression vectors of step e; g. culturing the mouse Sp2/0 cells of step f under conditions permitting expression and secretion of the immunoglobulin.