Transgenic non-human animals for producing chimeric antibodies

• US 7,501,552 B2
• Filed: 12/10/2004
• Issued: 03/10/2009
• Est. Priority Date: 08/28/1991
• Status: Expired due to Fees

• Current Assignee: Genpharm Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Medarex Incorporated (Bristol-Myers Squibb Company)
• Inventors: Kay, Robert M., Lonberg, Nils
• Primary Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US11/009,873
• Publication Number:
  US 20060015957A1
• Time in Patent Office: 1,551 Days
• Field of Search: 800/6, 800/18
• US Class Current: 800/6
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/462 Igs containing a variable region (Fv) from one specie and a constant region (Fc) from another
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

The invention relates to transgenic non-human animals capable of producing heterologous antibodies and methods for producing human sequence antibodies which bind to human antigens with substantial affinity.

Claims

1. A method for obtaining a somatically mutated antibody that binds to a predetermined antigen, which method comprises:

(a) immunizing a transgenic mouse with a predetermined antigen, wherein the transgenic mouse has a genome comprising inactivated endogenous murine heavy chain variable gene regions, and further comprising an unrearranged human heavy chain immunoglobulin variable region operably linked to a mu constant region gene segment,

wherein the unrearranged human heavy chain variable region comprises multiple human V_H gene segments, multiple human D gene segments, and multiple human J_H gene segments, and wherein the mu constant region is selected from the group consisting of (i) a human S-mu sequence and a human or mouse mu coding sequence and (ii) a mouse S-mu sequence and a human or mouse mu coding sequence,

(b) removing B lymphocytes from the transgenic mouse,

(c) identifying mouse IgG isotype immunoglobulin molecules that bind to the predetermined antigen expressed by the B lymphocytes; and

(d) isolating the IgG isotype immunoglobulin molecules that bind to the predetermined antigen, whereby a somatically mutated antibody that binds to the predetermined antigen is obtained from the B lymphocytes of step (c).

2. The method of claim 1, wherein the mouse IgG isotype immunoglobulin molecules are identified by screening cells derived from the B lymphocytes to determine which immunoglobulin isotype is expressed.

3. The method according to claim 1, wherein the IgG isotype immunoglobulin molecules are identified by isolating the nucleic acid molecules encoding the IgG immunoglobulin molecules from the B lymphocytes and determining the sequence of the heavy chain constant region.