×

Methods of modifying eukaryotic cells

  • US 6,596,541 B2
  • Filed: 02/16/2001
  • Issued: 07/22/2003
  • Est. Priority Date: 10/31/2000
  • Status: Expired due to Term
First Claim
Patent Images

1. A method of replacing, in whole or in part, in an isolated non-human eukaryotic cell, an endogenous immunoglobulin variable region gene locus with an homologous or orthologous human immunoglobulin variable gene locus comprising:

  • a) obtaining a large cloned genomic fragment greater than 20 kb containing, in whole or in part, the homologous or orthologous human immunoglobulin variable gene locus;

    b) using bacterial homologous recombination to genetically modify the cloned genomic fragment of (a) to create a large targeting vector for use in the eukaryotic cells (LTVEC), the LTVEC having homology arms totaling greater than 20 kb;

    c) introducing the LTVEC of (b) into the isolated eukaryotic cells to replace by homologous recombination, in whole or in part, the endogenous immunoglobulin variable gene locus; and

    d) using a quantitative assay to detect modification of allele (MOA) in the eukaryotic cells of (c) to identify those eukaryotic cells in which the endogenous immunoglobulin variable region gene locus has been replaced, in whole or in part, with the homologous or orthologous human immunoglobulin variable gene locus.

View all claims
  • 4 Assignments
Timeline View
Assignment View
    ×
    ×