Human anti-IFN-α antibodies
Provided are novel IFB-a binding molecules of human origin, particularly human-derived anti-IFN-α antibodies as well as IFN-α binding fragments, derivatives and variants thereof. In addition, pharmaceutical compositions, kits, and methods for use in diagnosis and therapy are described.
1. A human monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises in its variable regions the complementarily determining regions (CDRs) VH CDRs: amino acids 31-35 of SEQ ID NO:18 and amino acids 50-66 of SEQ ID NO: 18 and amino acids 99-115 of SEQ ID NO: 18; and VL CDRs: amino acids 24-35 of SEQ ID NO:20 and amino acids 51-57 of SEC) ID NO:20 and amino acids 90-100 of SEC) ID NO:20 and wherein the antibody has the following properties: (i) binds to human IFN-α subtypes IFNαI/13 (IFNαIb), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14 IFNα16, IFNα17 and IFNα21; and (ii) does not bind to IFN-ω, wherein the IFN-α antibody or IFN-α binding fragment thereof is labeled with a detectable label selected from the group consisting of an enzyme, a radioisotope, a fluorophore, a peptide and a heavy metal, or attached to a drug.
2. The monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 , comprising in its variable regions an amino acid sequence of the V H and V L region as depicted in SEQ ID NO:18 and 20, respectively.
3. The human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 , which binds an epitope of IFNα2 consisting of the amino acid sequence SAAWDETLLDKFYTELYQ (SEQ ID NO: 99) and/or RITLYLKEKKYSPCAWEV (SEQ ID NO: 100).
4. The human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 , which is an IgG1 isotype.
5. The human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 comprising a C H and/or C L constant region comprising an amino acid sequence selected from SEQ ID NOs.:72 and 74.
6. The human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 , which has an IC 50 value of ≤10 ng for at least 10 human IFN-α subtypes.
7. The human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 , which is selected from the group consisting of a single chain Fv fragment (scFv), an F(ab′) fragment, an F(ab) fragment, and an F(ab′)2 fragment and comprises in its variable regions the complementarity determining regions (CDRs)
V H CDRs: amino acids 31-35 of SEQ ID NO:18 and amino acids 50-66 of SEQ ID NO:18 and amino acids 99-115 of SEQ ID NO:18; and
V L CDRs: amino acids 24-35 of SEQ ID NO:20 and amino acids 51-57 of SEQ ID NO:20 and amino acids 90-100 of SEQ ID NO:20
and wherein the antibody has the following properties:
(i) binds to human IFN-α subtypes IFNα1/13 (IFNα1b), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14, IFNα16, IFNα17 and IFNα21;
and
(ii) does not bind to IFN-ω.
8. A method for diagnosing an inflammatory or autoimmune disease or condition associated with the expression of IFN-α in a subject comprising contacting a biological sample of the subject with the monoclonal anti-IFN-α antibody or IFN-α-binding fragment thereof of claim 1 , and detecting the presence of IFN-α in the biological sample.
9. A cDNA molecule encoding a monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof, wherein the antibody or IFN-α binding fragment thereof comprises in its variable regions the complementarity determining regions (CDRs)
V H CDRs: amino acids 31-35 of SEQ ID NO:18 and amino acids 50-66 of SEQ ID NO:18 and amino acids 99-115 of SEQ ID NO:18; and
V L CDRs: amino acids 24-35 of SEQ ID NO:20 and amino acids 51-57 of SEQ ID NO:20 and amino acids 90-100 of SEQ ID NO:20 and wherein the antibody has the following properties
(i) binds to human IFN-α subtypes IFNα1/13 (IFNα1b), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14, IFNα16, IFNα17 and IFNα21; and
(ii) does not bind to IFN-ω.
10. The cDNA molecule of claim 9 , wherein the monoclonal antibody or IFN-α binding fragment thereof binds an epitope of IFNα2 consisting of the amino acid sequence SAAWDETLLDKFYTELYQ (SEQ ID NO: 99) and/or RITLYLKEKKYSPCAWEV (SEQ ID NO: 100).
11. The cDNA molecule of claim 9 , wherein the monoclonal antibody or IFN-α binding fragment thereof comprises in its variable regions an amino acid sequence of the V H and V L region as depicted in SEQ ID NO:18 and 20, respectively.
12. The cDNA molecule of claim 9 , wherein the monoclonal antibody or IFN-α binding fragment thereof is an IgG1 isotype.
13. The cDNA molecule of claim 9 , wherein the monoclonal antibody or IFN-α binding fragment thereof comprises a C H and/or C L constant region comprising an amino acid sequence selected from SEQ ID NOs.:72 and 74.
14. The cDNA molecule of claim 9 , wherein the monoclonal antibody or IFN-α binding fragment thereof has an IC 50 value of ≤10 ng for at least 10 human IFN-α subtypes.
15. The cDNA molecule of claim 9 , wherein the monoclonal antibody or IFN-α binding fragment thereof is selected from the group consisting of a single chain Fv fragment (scFv), an F(ab′) fragment, an F(ab) fragment, and an F(ab′) 2 fragment wherein the antibody or IFN-α binding fragment thereof comprises in its variable regions the complementarity determining regions (CDRs)
V H CDRs: amino acids 31-35 of SEQ ID NO:18 and amino acids 50-66 of SEQ ID NO:18 and amino acids 99-115 of SEQ ID NO:18; and
V L CDRs: amino acids 24-35 of SEQ ID NO:20 and amino acids 51-57 of SEQ ID NO:20 and amino acids 90-100 of SEQ ID NO:20 and wherein the antibody has the following properties
(i) binds to human IFN-α subtypes IFNα1/13 (IFNα1b), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14, IFNα16, IFNα17 and IFNα21; and
(ii) does not bind to IFN-ω.
16. The cDNA molecule of claim 9 , encoding the variable region and at least part of the constant domain.
17. The cDNA molecule of claim 9 , wherein the monoclonal antibody or IFN-α binding fragment thereof is a human monoclonal antibody or IFN-α binding fragment thereof.
18. A vector comprising the cDNA molecule of claim 9 .
19. The vector of claim 18 , wherein the vector is an expression vector.
20. An isolated host cell comprising the vector of claim 19 .
21. The host cell of claim 20 , which is a prokaryotic or eukaryotic host cell.
22. A method of producing a monoclonal anti-IFN-α antibody or IFN-α binding fragment thereof encoded by the vector in the host cell of claim 20 , comprising:
(a) culturing in a cell culture the host cell under conditions allowing the expression of the antibody encoded by the cDNA molecule; and
(b) isolating the antibody from the cell culture.
23. A polynucleotide linked to a heterologous nucleic acid, wherein the polynucleotide is selected from the group consisting of:
(a) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising the complementarity determining regions (CDRs) of the VH variable region amino acid sequence depicted in SEQ ID NO: 18, wherein the immunoglobulin heavy chain VH when paired with a light chain variable region (VL) comprising the amino acid sequence depicted in SEQ ID NO: 20
(i) binds to human IFN-α subtypes IFNα1/13 (IFNα1b), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14, IFNα16, IFNα17 and IFNα21; and
(ii) does not bind to IFN-ω;
(b) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising the complementarity determining regions (CDRs) of the VL variable region amino acid sequence depicted in SEQ ID NO: 20, wherein the immunoglobulin light chain VL when paired with a heavy chain variable region (VH) comprising the amino acid sequence depicted in SEQ ID NO: 18
i) binds to human IFN-α subtypes IFNα1/13 (IFNα1b), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14, IFNα16, IFNα17 and IFNα21; and
(ii) does not bind to IFN-ω;
(c) a polynucleotide encoding
(i) an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising the complementarity determining regions (CDRs) of the VH variable region amino acid sequence depicted in SEQ ID NO: 18; and
(ii) an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising the complementarity determining regions (CDRs) of the VL variable region amino acid sequence depicted in SEQ ID NO: 20;
(d) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising the amino acid sequence of SEQ ID NO: 18;
(e) a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising the amino acid sequence of SEQ ID NO: 20; and
(f) a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising the amino acid sequence of SEQ ID NO: 18 and an immunoglobulin light chain or a fragment thereof comprising the amino acid sequence of SEQ ID NO: 20.
24. The polynucleotide of claim 23 , wherein the polynucleotide further encodes an amino acid sequence of a C H and/or C L constant region comprising an amino acid sequence selected from SEQ ID NOs.:72 and 74.
25. The polynucleotide of claim 23 , wherein the heterologous nucleic acid is a regulatory element.
26. The polynucleotide of claim 25 , wherein the regulatory element is a promoter, an enhancer, a transcription termination sequence or a leader sequence.
27. A vector comprising one or more of polynucleotides of claim 23 .
28. The vector of claim 27 , wherein the vector is an expression vector.
29. An isolated host cell comprising one or more vectors of claim 28 .
30. The host cell of claim 29 , which is a prokaryotic or eukaryotic host cell.
31. A method of producing a monoclonal antibody that binds to human IFN-α subtypes IFNα1/13 (IFNα1b), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14, IFNα16, IFNα17 and IFNα21; and does not bind to IFN-ω or an immunoglobulin chain thereof encoded by the vector in the host cell of claim 29 , which comprises:
(a) culturing in a cell culture the host cell under conditions allowing the expression of the antibody or immunoglobulin chain; and
(b) isolating the antibody or immunoglobulin chain from the cell culture.
32. The method of claim 31 , wherein the monoclonal antibody is a human antibody.
33. A method of treating an inflammatory or autoimmune disease or condition associated with the expression of IFN-α comprising administering to a subject in need of such treatment an effective amount of a human anti-IFN-α antibody or IFN-α binding fragment thereof produced by the method of claim 22 or 31 .
34. A method of treating an inflammatory or autoimmune disease or condition associated with the expression of IFN-α comprising administering to a subject in need of such treatment an effective amount of a human anti-IFN-α antibody or IFN-α binding fragment thereof encoded by the cDNA molecule of claim 9 or the polynucleotides of claim 23 .