Bispecific antigen binding proteins

• US 10,640,555 B2
• Filed: 04/24/2017
• Issued: 05/05/2020
• Est. Priority Date: 06/16/2009
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Imhof-Jung, Sabine, Klein, Christian, Regula, Joerg Thomas, Schaefer, Wolfgang, Schanzer, Juergen Michael
• Primary Examiner(s): Natarajan, Meera
• Application Number:
  US15/495,641
• Publication Number:
  US 20180079805A1
• Time in Patent Office: 1,107 Days
• Field of Search: None
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/64 comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/66 comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2319/00 Fusion polypeptide

Abstract

The present invention relates to bispecific antigen binding proteins, methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

Claims

1. A method for the preparation of a bispecific antigen binding protein comprising:

a) two light chains and two heavy chains of an antibody that comprises two Fab fragments and that specifically binds to a first antigen; and

b) two additional Fab fragments of an antibody which specifically binds to a second antigen, wherein the additional Fab fragments are both fused via a peptide connector either at the C- or N-termini of the heavy chains of a);

wherein the bispecific antigen binding protein also comprises a structural modification selected from the group consisting of;

i) in both Fab fragments of a) or in both Fab fragments of b) the variable domains VL and VH are replaced by each other, and the constant domains CL and CH1 are replaced by each other, or the constant domains CL and CH1 are replaced by each other;

ii) in both Fab fragments of a)

the variable domains VL and VH are replaced by each other,

and

the constant domains CL and CH1 are replaced by each other, and in both Fab fragments of b)

the variable domains VL and VH are replaced by each other,

or

the constant domains CL and CH1 are replaced by each other;

iii) in both Fab fragments of a)

the variable domains VL and VH are replaced by each other,

or

the constant domains CL and CH1 are replaced by each other,

and

in both Fab fragments of b)

the variable domains VL and VH are replaced by each other,

and

the constant domains CL and CH1 are replaced by each other;

iv) in both Fab fragments of a)

the variable domains VL and VH are replaced by each other,

and

in both Fab fragments of b)

the constant domains CL and CH1 are replaced by each other;

and

v) in both Fab fragments of a)

the constant domains CL and CH1 are replaced by each other,

and

in both Fab fragments of b)

the variable domains VL and VH are replaced by each other,

comprising the steps of

a) transforming a host cell with vectors comprising nucleic acid molecules encoding said bispecific antigen binding protein;

b) culturing the host cell under conditions that allow synthesis of said antigen binding protein molecule; and

c) recovering said antigen binding protein molecule from said culture.

2. A host cell comprising the vectors according to claim 1.