US 2004/0167319 A1 · Human monoclonal antibodies against CD20

Abstract

Isolated human monoclonal antibodies which bind to and inhibit human CD20, and related antibody-based compositions and molecules, are disclosed. The human antibodies can be produced by a transfectoma or in a non-human transgenic animal, e.g., a transgenic mouse, capable of producing multiple isotypes of human monoclonal antibodies by undergoing V-D-J recombination and isotype switching. Also disclosed are pharmaceutical compositions comprising the human antibodies, non-human transgenic animals and hybridomas which produce the human antibodies, and therapeutic and diagnostic methods for using the human antibodies.

Claims102

Dependent claims

An isolated human monoclonal antibody which binds to human CD20.

An isolated human monoclonal antibody which binds to an epitope on CD20

(i) which does not comprise or require the amino acid residue proline at position 172;
(ii) which does not comprise or require the amino acid residues alanine at position 170 or proline at position 172;
(iii) which comprises or requires the amino acid residues asparagine at position 163 and asparagine at position 166;
(iv) which does not comprise or require the amino acid residue proline at position 172, but which comprises or requires the amino acid residues asparagine at position 163 and asparagine at position 166;
or
(v) which does not comprise or require the amino acid residues alanine at position 170 or proline at position 172, but which comprises or requires the amino acid residues asparagine at position 163 and asparagine at position 166.

3 more elements

An isolated human monoclonal antibody which binds to CD20, wherein the antibody has one or more of the following characteristics:

(i) binds to mutant P172S CD20 (proline at position 172 mutated to serine) with at least the same affinity as to human CD20;
(ii) binds to mutant A×P (alanine at position 170 mutated to serine, and proline at position 172 mutated to serine) with at least the same affinity as to human CD20;
(iii) shows a reduced binding of 50% or more to mutant N166D (asparagine at position 166 mutated to aspartic acid) compared to human CD20 at an antibody concentration of 10 μg/ml;
or
(iv) shows a reduced binding of 50% or more to mutant N163D (asparagine at position 163 mutated to aspartic acid) compared to human CD20 at an antibody concentration of 10 μg/ml.

2 more elements

An isolated human monoclonal antibody which binds to an epitope in the small first extracellular loop of human CD20.

An isolated human monoclonal antibody which binds to a discontinuous epitope on CD20.

An isolated human monoclonal antibody which binds to a discontinuous epitope on CD20, wherein the epitope comprises part of the first small extracellular loop and part of the second extracellular loop.

An isolated human monoclonal antibody which binds to a discontinuous epitope on CD20, wherein the epitope has residues AGIYAP of the small first extracellular loop and residues MESLNFIRAHTPYI of the second extracellular loop.

An isolated human monoclonal antibody which binds to human CD20 comprising at least one CDR sequence selected from the group consisting of:

(i) SEQ ID NOs:
13, 14, 15, 16, 17, or 18;
(ii) conservative sequence modifications of the sequences listed in (i);
and
(iii) fragments of any one of the sequences defined in (i) or (ii) which retain the ability to bind to human CD20.

2 more elements

An isolated human monoclonal antibody which binds to human CD20 comprising at least one CDR sequence selected from the group consisting of:

(i) SEQ ID NOs:
19, 20, 21, 22, 23, or 24;
(ii) conservative sequence modifications of the sequences listed in (i);
and
(iii) fragments of any one of the sequences defined in (i) or (ii) which retain the ability to bind to human CD20.

2 more elements

An isolated human monoclonal antibody which binds to human CD20 comprising at least one CDR sequence selected from the group consisting of:

(i) SEQ ID NOs:
25, 26, 27, 28, 29, or 30;
(ii) conservative sequence modifications of the sequences listed in (i);
and
(iii) fragments of any one of the sequences defined in (i) or (ii) which retain the ability to bind to human CD20.

2 more elements

A hybridoma which produces a human monoclonal antibody encoded by human IgG heavy chain and human kappa light chain nucleic acids comprising nucleotide sequences in their variable regions as set forth in SEQ ID NOs:1, 5, or 9 and SEQ ID NOs:3, 7, or 11, respectively, and conservative sequence modifications thereof.

A hybridoma which produces a human monoclonal having IgG heavy chain and kappa light chain variable regions which comprise the amino acid sequences as set forth in SEQ ID NOs:2, 6, or 10 and SEQ ID NOs:

4, 8, or 12, respectively, and conservative sequence modifications thereof.

A transfectoma which produces a human monoclonal antibody encoded by human IgG heavy chain and human kappa light chain nucleic acids comprising nucleotide sequences in their variable regions as set forth in SEQ ID NOs:1, 5, or 9 and SEQ ID NOs:3, 7, or 11, respectively, and conservative sequence modifications thereof.

A transfectoma which produces a human monoclonal antibody having IgG heavy chain and kappa light chain variable regions which comprise the amino acid sequences as set forth in SEQ ID NOs:2, 6, or 10 and SEQ ID NOs:4, 8, or 12, respectively, and conservative sequence modifications thereof.

A eukaryotic or prokaryotic host cell which produces a human monoclonal antibody having heavy chain and light chain variable regions which comprise the amino acid sequences as set forth in SEQ ID NOs:2, 6, or 10 and SEQ ID NOs:4, 8, or 12, respectively, and conservative sequence modifications thereof.

A transgenic non-human animal or plant which produces a human monoclonal antibody having heavy chain and light chain variable regions which comprise the amino acid sequences as set forth in SEQ ID NOs:2, 6, or 10 and SEQ ID NOs:4, 8, or 12, respectively, and conservative sequence modifications thereof.

A method of producing a human monoclonal antibody which binds to human CD20, comprising:

immunizing a transgenic non-human animal having a genome comprising a human heavy chain transgene and a human light chain transgene with human CD20 or a cell expressing human CD20, such that antibodies are produced by B cells of the animal;
isolating B cells of the animal;
fusing the B cells with myeloma cells to form immortal, hybridoma cells that secrete human monoclonal antibodies specific for human CD20;
and
isolating the human monoclonal antibodies specific for CD20 from the culture supernatant of the hybridoma, or the transfectoma derived from such hybridoma.

2 more elements

A human monoclonal antibody which binds to human CD20 obtained by:

immunizing a transgenic non-human animal having a genome comprising a human heavy chain transgene and a human light chain transgene with a cell which has been transfected with human CD20, such that antibodies are produced by B cells of the animal;
isolating B cells of the animal;
fusing the B cells with myeloma cells to form immortal, hybridoma cells that secrete human monoclonal antibodies specific for human CD20;
and
isolating the human monoclonal antibodies specific for CD20 from the culture supernatant of the hybridoma, or the transfectoma derived from such hybridoma.

2 more elements

An isolated human antibody comprising a heavy chain variable region amino acid sequence derived from a human VH3-13/DP-44 germline sequence (SEQ ID NO:54) and a light chain variable region amino acid sequence derived from a human VL-L6/JK4-CK (SEQ ID NO:55) germline sequence, wherein the human antibody binds to human CD20.

An isolated human antibody comprising a heavy chain variable region amino acid sequence derived from a human VH3-09/JH6b germline sequence (SEQ ID NO:56) and a light chain variable region amino acid sequence derived from a human VL-L6/JK5 germline sequence (SEQ ID NO:57), wherein the human antibody binds to human CD20.

A composition comprising a first human antibody and a second human antibody, both of which bind to human CD20, wherein the first antibody has a human heavy chain and human kappa light chain variable regions comprising the amino acid sequences as set forth in SEQ ID NO:2 and SEQ ID NO:4, respectively, and conservative sequence modifications thereof, and wherein the second antibody has a human heavy chain and human kappa light chain variable regions comprising the amino acid sequences as set forth in SEQ ID NO:10 and SEQ ID NO:12, respectively, and conservative sequence modifications thereof.

An expression vector comprising a nucleotide sequence encoding the variable region of a light chain, heavy chain or both light and heavy chains of a human antibody which binds human CD20.

An expression vector comprising a nucleotide sequence encoding a heavy chain variable region comprising a nucleotide selected from the group consisting of the nucleotide sequences as set forth in SEQ ID NOs:

1, 5, and 9, and a light variable region comprising a nucleotide sequence selected from the group consisting of the nucleotide sequences as set forth in SEQ ID NOs:
3, 7, and 11, and conservative modifications thereof.

An expression vector comprising a nucleotide sequence encoding a heavy chain variable region comprising an amino acid sequence selected from the group consisting of the amino acid sequences as set forth in SEQ ID NOs:2, 6, and 10, and a light chain variable region comprising the amino acid sequence selected from the group consisting of the amino acid sequences as set forth in shown in SEQ ID NOs:

4, 8, and 12, and conservative sequence modifications thereof.

An anti-idiotypic antibody binding to 2F2, 11B8 or 7D8.

Dependencies are inferred from the claim text.

Human monoclonal antibodies against CD20

Life of this patent

Human monoclonal antibodies against CD20

Life of this patent

Abstract

Claims102