US 2017/0362323 A1 · HETERODIMER PROTEIN COMPOSITION

Abstract

According to the present invention, a heterodimer protein composition that is composed of a first polypeptide comprising an immunoglobulin heavy chain constant region and a second polypeptide comprising CL-Fc prepared by fusion of an immunoglobulin light chain constant region (CL) and Fc region, and also has a deletion or substitution of Cys residues involved in disulfide bonds between CL and CH1, a purification method of the protein composition thereof, a preparation method of the protein composition thereof, a DNA and a vector encoding the protein composition thereof are provided

Claims40

Dependent claims

(canceled)

A DNA encoding a heterodimer protein, the heterodimer protein being

(i) a heterodimer protein composed of
a first polypeptide comprising a heavy chain variable region (VH) and an immunoglobulin G (IgG)1 heavy chain constant region (CH), and
a second polypeptide comprising a light chain variable region (VL) and a fusion polypeptide (CL-Fc) prepared by fusion of a light chain constant region (CL) and an Fc region of IgG1,
wherein the heterodimer protein has a modification selected from the group consisting of:
(a-i) substitutions of C220S in the first polypeptide and H435R in the second polypeptide,
(b-i) substitutions of C214S and H435R in the second polypeptide,
(c-i) substitutions of C220S in the first polypeptide and C214S and H435R in the second polypeptide,
(d-i) substitutions of C220S in the first polypeptide and C220S, C214S and H435R in the second polypeptide,
(e-i) substitutions of C220S in the first polypeptide and C214S and H435R in the second polypeptide and a deletion of positions 216-220 in the second polypeptide,
(f-i) substitutions of C220S in the first polypeptide and C214S, H435R and Y436F in the second polypeptide,
(g-i) substitutions of C220S in the first polypeptide and C214S, H435R and Y436F in the second polypeptide and a deletion of positions 216-220 in the second polypeptide,
(h-i) substitutions of C220S, K332A and P331S in the first polypeptide, and C214S, K322A, P331S, H435R and Y436F in the second polypeptide and a deletion of positions 216-220 in the second polypeptide, and
(j-i) substitutions of C220S and I332E in the first polypeptide and C214S, I332E, H435R and Y436F in the second polypeptide and a deletion of positions 216-220 in the second polypeptide, numbering according to the EU index of Kabat (“the EU index”), or
(ii) a heterodimer protein composed of
a first polypeptide comprising a VH and an IgG4 CH, and
a second polypeptide comprising a VL and a CL-Fc prepared by fusion of a CL and an Fc region of IgG4,
wherein the heterodimer protein has a modification selected from the group consisting of:
(a-ii) substitutions of C131S in the first polypeptide and C214S and H435R in the second polypeptide,
(b-ii) substitutions of C131S in the first polypeptide and C214S, H435R and Y436F in the second polypeptide,
(c-ii) substitutions of C131S and R409K in the first polypeptide and C214S, R409K, H435R and Y436F in the second polypeptide,
(d-ii) substitutions of C131S, S228P and R409K in the first polypeptide and C214S, S228P, R409K, H435R and Y436F in the second polypeptide,
(e-ii) substitutions of C131S, S228P, L235E and R409K in the first polypeptide and C214S, S228P, L235E, R409K, H435R and Y436F in the second polypeptide,
(f-ii) substitutions of C131S and R133K in the first polypeptide and C214S and H435R in the second polypeptide,
(g-ii) substitutions of C131S and R133K in the first polypeptide and C214S, H435R and Y436F in the second polypeptide,
(h-ii) substitutions of C131S, R133K and R409K in the first polypeptide and C214S, R409K, H435R and Y436F in the second polypeptide,
(j-ii) substitutions of C131S, R133K, S228P and R409K in the first polypeptide and C214S, S228P, R409K, H435R and Y436F in the second polypeptide, and
(k-ii) substitutions of C131S, R133K, S228P, L235E and R409K in the first polypeptide and C214S, S228P, L235E, R409K, H435R and Y436F in the second polypeptide, numbering according to the EU index of Kabat,
wherein in (i) and (ii), the VH of the first polypeptide and the VL of the second polypeptide forms a binding domain.

25 more elements

A DNA encoding a heterodimer protein, the heterodimer protein being

(i) a heterodimer protein composed of
a first polypeptide comprising a light chain variable region (VL) and an IgG1 heavy chain constant region (CH), and
a second polypeptide comprising a heavy chain variable region (VH) and a fusion polypeptide (CL-Fc) prepared by fusion of a light chain constant region (CL) and an Fc region of IgG1,
wherein the heterodimer protein has a modification selected from the group consisting of:
(a-i) substitutions of C220S in the first polypeptide and H435R in the second polypeptide,
(b-i) substitutions of C214S and H435R in the second polypeptide,
(c-i) substitutions of C220S in the first polypeptide and C214S and H435R in the second polypeptide,
(d-i) substitutions of C220S in the first polypeptide and C220S, C214S and H435R in the second polypeptide,
(e-i) substitutions of C220S in the first polypeptide and C214S and H435R in the second polypeptide and a deletion of positions 216-220 in the second polypeptide,
(f-i) substitutions of C220S in the first polypeptide and C214S, H435R and Y436F in the second polypeptide,
(g-i) substitutions of C220S in the first polypeptide and C214S, H435R and Y436F in the second polypeptide and a deletion of positions 216-220 in the second polypeptide,
(h-i) substitutions of C220S, K332A and P331S in the first polypeptide, and C214S, K322A, P331S, H435R and Y436F in the second polypeptide and a deletion of positions 216-220 in the second polypeptide, and
(j-i) substitutions of C220S and I332E in the first polypeptide and C214S, I332E, H435R and Y436F in the second polypeptide and a deletion of positions 216-220 in the second polypeptide, numbering according to the EU index of Kabat (“the EU index”), or
(ii) a heterodimer protein composed of
a first polypeptide comprising a VL and an IgG4 CH, and
a second polypeptide comprising a VH and a CL-Fc prepared by fusion of a CL and an Fc region of IgG4,
wherein the heterodimer protein has a modification selected from the group consisting of:
(a-ii) substitutions of C131S in the first polypeptide and C214S and H435R in the second polypeptide,
(b-ii) substitutions of C131S in the first polypeptide and C214S, H435R and Y436F in the second polypeptide,
(c-ii) substitutions of C131S and R409K in the first polypeptide and C214S, R409K, H435R and Y436F in the second polypeptide,
(d-ii) substitutions of C131S, S228P and R409K in the first polypeptide and C214S, S228P, R409K, H435R and Y436F in the second polypeptide,
(e-ii) substitutions of C131S, S228P, L235E and R409K in the first polypeptide and C214S, S228P, L235E, R409K, H435R and Y436F in the second polypeptide,
(f-ii) substitutions of C131S and R133K in the first polypeptide and C214S and H435R in the second polypeptide,
(g-ii) substitutions of C131S and R133K in the first polypeptide and C214S, H435R and Y436F in the second polypeptide,
(h-ii) substitutions of C131S, R133K and R409K in the first polypeptide and C214S, R409K, H435R and Y436F in the second polypeptide,
(j-ii) substitutions of C131S, R133K, S228P and R409K in the first polypeptide and C214S, S228P, R409K, H435R and Y436F in the second polypeptide and
(k-ii) substitutions of C131S, R133K, S228P, L235E and R409K in the first polypeptide and C214S, S228P, L235E, R409K, H435R and Y436F in the second polypeptide, numbering according to the EU index of Kabat,
wherein in (i) and (ii), the VL of the first polypeptide and the VH of the second polypeptide forms a binding domain.

25 more elements

Dependencies are inferred from the claim text.

Facts

Application

15/624,581Filed 15 Jun 2017

Priority

5 Oct 2012

Published

21 Dec 2017

Term ends

Not estimatedThe source records don’t support an estimate.

Classification10 codes

C07K 16/2833
Against the immunoglobulin superfamily: against MHC-molecules, e.g. HLA-molecules
C07K 16/2887
Against receptors, cell surface antigens or cell surface determinants: against CD20
C07K 16/32
Against material from animals or humans: against translation products of oncogenes
C07K 16/468
Hybrid immunoglobulins: immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies

Family

Application61/710,221Us provisional application · filed 5 Oct 2012

Application14/046,478Division · filed 4 Oct 2013

Application15/624,581Division

US 10,494,437 B2Related patent

US 2014/0120581 A1Published application

US 9,714,291 B2Related patent

US 2017/0362323 A1This patent

From the US filing history. Foreign family members aren’t listed.

Heterodimer protein composition

Life of this patent

Abstract

Claims40

Heterodimer protein composition

Life of this patent

Abstract

Claims40