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 recombinant human monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof, wherein the antibody or IFN-α-binding fragment thereof comprises:
a variable heavy (VH) chain sequence comprising the
sequence of
(SEQ ID NO: 18)
EVQLLESGAEVKRPGSSVRVSCRASGDTFSSYPISWVRQAPGQGLEWMGR
ILPALGVTNYAQNFRGRITITADKSPLTAYLELSSLRFEDTAVYYCASPS
ADIIPSILGTTLFAFWGQGSLVTVSS
and
a variable light (VL) chain sequence comprising the
sequence of
(SEQ ID NO: 20)
EIVLTQSPGTLSLSPGEGATLSCRASQNVSRHYLTWYQQKPGQSPRLLIY
GGSSRAIGVPDRFSGGGSGTDFTLTISRLEPEDFAVFYCQSYHSPPPVYT
FGQGTKVEIK,
wherein the antibody has the following properties:
(i) binds to human IFN-α subtypes IFNα1/13(IFNαlb), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNα10, IFNα14, IFNα16, IFNα17 and IFNα21; and
(ii) does not bind to IFN-ω.
2. The recombinant human monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof of claim 1 , wherein amino acids 1-6 and 121-126 of (SEQ ID NO: 18) have not been isolated or sequenced from a human.
3. The recombinant human monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof of claim 2 , wherein amino acids 1-6 and 121-126 of (SEQ ID NO: 18) are obtained from a database.
4. The recombinant human monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof of claim 1 , wherein amino acids 1-6 and 105-110 of (SEQ ID NO: 20) have not been isolated or sequenced from a human.
5. The recombinant human monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof of claim 4 , wherein amino acids 1-6 and 105-110 (SEQ ID NO: 20) are obtained from a database.
6. The recombinant human monoclonal anti-interferon-alpha (IFN-α) antibody or an IFN-α binding fragment thereof of claim 1 , which is an IgG1 isotype.
7. The recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 , further comprising a C H and/or C L constant region comprising an amino acid sequence selected from SEQ ID NOs.:72 and 74.
8. The recombinant 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.
9. A composition comprising the recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 1 .
10. The composition of claim 9 , further comprising a pharmaceutically acceptable carrier.
11. A method of producing the recombinant human monoclonal IFN-α antibody of claim 1 comprising the steps of:
(a) culturing in a cell culture a host cell comprising a vector comprising a cDNA molecule or a polynucleotide encoding
a variable heavy (VH) chain sequence comprising the
sequence of
(SEQ ID NO: 18)
EVQLLESGAEVKRPGSSVRVSCRASGDTFSSYPISWVRQAPGQGLEWMGR
ILPALGVTNYAQNFRGRITITADKSPLTAYLELSSLRFEDTAVYYCASPS
ADIIPSILGTTLFAFWGQGSLVTVSS
and
a variable light (VL) chain sequence comprising the
sequence of
(SEQ ID NO: 20)
EIVLTQSPGTLSLSPGEGATLSCRASQNVSRHYLTWYQQKPGQSPRLLIY
GGSSRATGVPDRFSGGGSGTDFTLTISRLEPEDFAVFYCQSYHSPPPVYT
FGQGTKVEIK,
under conditions allowing the expression of the antibody encoded by the cDNA molecule or the polynucleotide; and
(b) isolating the antibody from the cell culture.
12. A method of treating an inflammatory or autoimmune disease or condition associated with the expression of IFN-α comprising administering to a subject a therapeutically effective amount of the recombinant human monoclonal anti-IFN-α antibody or IFN-α binding fragment thereof of claim 1 .
13. The method of claim 12 , wherein the method comprises administering to the subject a therapeutically effective dose of the recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof.
14. The method of claim 12 , wherein the recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof is administered intravenously.
15. The method of claim 14 , wherein the recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof is administered at a dose of between 1 μg to 10 mg per day.
16. The method of claim 15 , wherein the recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof is administered by an infusion.
17. The method of claim 16 , wherein the recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof is administered by infusion at a dose of between 1 1tg to 10 mg units per kilogram of body weight per minute.
18. A method of treating an inflammatory or autoimmune disease or condition associated with the expression of IFN-α comprising administering to a subject a therapeutically effective amount of a composition comprising the recombinant human monoclonal anti-IFN-α antibody or IFN-α-binding fragment thereof of claim 1 .
19. The method of claim 18 , wherein the method comprises administering to the subject a therapeutically effective dose of the composition.
20. The method of claim 18 , wherein the composition is administered intravenously.
21. The method of claim 20 , wherein composition is administered at a dose of between 1 μg to 10 mg units per day.
22. The method of claim 21 , wherein the composition is administered by infusion.
23. The method of claim 22 , wherein the composition is administered by infusion at a dose of between 1 μg to 10 mg units per kilogram of body weight per minute.
24. A recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof produced according to a method comprising the steps of:
(a) culturing a plurality of B cells obtained from a subject having autoimmune polyendocrinopathy syndrome type 1 (ASP1) or autoimmune polyendocrinopathy-candidiasis ectodermal dystrophy (APECED) under conditions allowing only a definite life span of the B cells;
(b) identifying a B cell reactive to human IFN-α subtypes IFNα1/13 (IFNαlb), IFNα2, IFNα4, IFNα5, IFNα6, IFNα7, IFNα8, IFNαl 0, IFNα14, IFNα16, IFNα17 and IFNα21 and not reactive to IFN-ω;
(c) sequencing an antibody produced from the B cell; and
(d) cloning the antibody of (c) to produce a recombinant antibody using one or more primers, wherein the one or more primers introduce a sequence variation into the recombinant antibody compared to the sequence of (c),
wherein the IFN-α antibody or IFN-α-binding fragment thereof comprises:
a variable heavy (VH) chain sequence comprising the
sequence of
(SEQ ID NO: 18)
EVQLLESGAEVKRPGSSVRVSCRASGDTFSSYPISWVRQAPGQGLEWMGR
ILPALGVTNYAQNFRGRITITADKSPLTAYLELSSLRFEDTAVYYCASPS
ADIIPSILGTTLFAFWGQGSLVTVSS
and
a variable light (VL) chain sequence comprising the
sequence of
(SEQ ID NO: 20)
EIVLTQSPGTLSLSPGEGATLSCRASQNVSRHYLTWYQQKPGQSPRLLIY
GGSSRATGVPDRFSGGGSGTDFTLTISRLEPEDFAVFYCQSYHSPPPVY
TFGQGTKVEIK.
25. The recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 24 , wherein the conditions allowing only a definite life span of the B cells comprise exposing the plurality of B-cells to Epstein-Barr Virus (EBV) supernatant to produce a stimulated plurality of B cells.
26. The recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 25 , wherein the conditions allowing only a definite life span of the B cells comprise exposing the plurality of stimulated B-cells to CpG to produce an activated plurality of B cells.
27. The recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 26 , wherein the conditions allowing only a definite life span of the B cells further comprise culturing the plurality of stimulated B-cells under oligoclonal conditions with an irradiated feeder cell population from a heathy donor.
28. The recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 27 , wherein the irradiated feeder cell population comprises peripheral blood mononuclear cells (PMBCs).
29. The recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 24 , wherein a framework region of a VH sequence encoding the recombinant antibody comprises a sequence variation when compared to a VH sequence of an antibody isolated from human serum.
30. The recombinant human monoclonal IFN-α antibody or IFN-α binding fragment thereof of claim 29 , wherein a framework region of a VL sequence encoding the recombinant antibody comprises a sequence variation when compared to a VL sequence of an antibody isolated from human serum.