ANTI-DLL4 ANTIBODIES AND USES THEREOF

• US 20110318339A1
• Filed: 11/04/2009
• Published: 12/29/2011
• Est. Priority Date: 11/07/2008
• Status: Active Grant

• Current Assignee: Taurus Biosciences LLC (Ligand Pharmaceuticals, Inc.)
• Original Assignee: Fabrus, LLC. (Eloxx Pharmaceuticals, Inc.)
• Inventors: Smider, Vaughn, Mao, Helen Hongyuan, Bentley, Cornelia, Chase, Tyson
• Granted Patent: US 9,403,904 B2
• Time in Patent Office: Days
• US Class Current: 424/133.1
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 16/2884 against CD44
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/14 Specific host cells or culture conditions, e.g. components, pH or temperature
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/51 Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• 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 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/74 : Inducing cell proliferation
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 15/1037 : Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C40B 40/08 : Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/08 : Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support
• CPC Class Codes: G01N 33/6845 : Methods of identifying protein-protein interactions in protein mixtures

Abstract

Provided herein are anti-DLL4 antibodies and methods of using anti-DLL4 antibodies as therapeutic agents in diseases or disorders associated with DLL4.

Claims

1-135. -135. (canceled)

136. An isolated antibody or antigen-binding fragment thereof that specifically binds to human DLL4, wherein the antibody comprises at least one at least one heavy chain variable domain and at least one light chain variable domain, wherein each heavy chain variable domain comprises a complementary determining region (CDR) selected from the group consisting of CDRH1, CDRH2, and CDRH3 and each light chain variable domain comprises a CDR selected from the group consisting of CDRL1, CDRL2, and CDRL3, wherein:

a) CDRH3 has an amino acid sequence selected from the group consisting of EEYSSSSAEYFQH (SEQ ID NO;

851), RGYSYGYDYFDY (SEQ ID NO;

852), EYYDFWSGYYTDYFDY (SEQ ID NO;

853), EGYSSSWYDYFDY (SEQ ID NO;

854), ANWGDYFDY (SEQ ID NO;

855), DDYGGNSDYFDY (SEQ ID NO;

856), EGYCSGGSCYS (SEQ ID NO;

857), EYYYGSGSYYNDYFDY (SEQ ID NO;

858), GCYCSSTSCYADYYYYYGMDV (SEQ ID NO;

859), GSCYSYWYFDL (SEQ ID NO;

860), and an amino acid sequence that exhibits at least about 65% sequence identity to an amino acid sequence of SEQ ID NOS;

851-860;

b) CDRH2 has an amino acid sequence selected from the group consisting of IINPSGGSTSYAQKFQG (SEQ ID NO;

844), IIYPGDSDTRYSPSFQG (SEQ ID NO;

845), RTYYRSKWYNDYAVSVKS (SEQ ID NO;

846), EINHSGSTNYNPSLKS (SEQ ID NO;

847), INSNAGNGNTKYSQEFQG (SEQ ID NO;

     848), WMNPNSGNTGYAQKFQG (SEQ ID NO;

849), YIYYSGSTYYNPSLKS (SEQ ID NO;

850), and an amino acid sequence that exhibits at least about 65% sequence identity to an amino acid sequence of SEQ ID NOS;

844-850;

c) CDRH1 has an amino acid sequence selected from the group consisting of GYTFTSYYMH (SEQ ID NO;

     830), GYSFTSYWIG (SEQ ID NO;

831), GDSVSSNSAA (SEQ ID NO;

832), GGSFSGYYWS (SEQ ID NO;

833), GYTFTSYAMH (SEQ ID NO;

834), GYTFTSYAIN (SEQ ID NO;

835), GGSISSGGYY (SEQ ID NO;

836), a sequence of amino acids that exhibits at least 65% sequence identity to a sequence of amino acids set forth in any of SEQ ID NOS;

831-836, with the proviso that the sequence of amino acids is not GYTFTSYVIN (SEQ ID NO;

904), and an amino acid sequence that exhibits at least about 72% sequence identity to an amino acid sequence of SEQ ID NO;

830;

d) CDRL3 has an amino acid sequence selected from the group consisting of QQRSNWPPWT (SEQ ID NO;

881), VLYMGSGISYV (SEQ ID NO;

882), MIWHSSASFV (SEQ ID NO;

     883), QQYNNWPPWT (SEQ ID NO;

     884), QANSFPPWT (SEQ ID NO;

885), QQYGSSPPWT (SEQ ID NO;

     886), QQYNSYSPWT (SEQ ID NO;

887), MQRIEFPSWT (SEQ ID NO;

     888), SSYTSSSTLFV (SEQ ID NO;

     889), QVYESSANFV (SEQ ID NO;

     890), and an amino acid sequence that exhibits at least about 65% sequence identity to an amino acid sequence of SEQ ID NOS;

881-890;

e) CDRL2 has an amino acid sequence selected from the group consisting of DASNRAT (SEQ ID NO;

871), STNTRSS (SEQ ID NO;

     872), YYSDSSK (SEQ ID NO;

873), GASTRAT (SEQ ID NO;

874), AASSLQS (SEQ ID NO;

875), GASSRAT (SEQ ID NO;

876), DASSLGS (SEQ ID NO;

877), TLSYRAS (SEQ ID NO;

878), EVSNRPS (SEQ ID NO;

879), HYSDSDK (SEQ ID NO;

880), and an amino acid sequence that exhibits at least about 65% sequence identity to an amino acid sequence of SEQ ID NOS;

871-880; and

f) CDRL1 has an amino acid sequence selected from the group consisting of RASQSVSSYLA (SEQ ID NO;

     861), GLSSGSVSTSYYPS (SEQ ID NO;

862), TLRSGINLGSYRIF (SEQ ID NO;

863), RASQSVSSNLA (SEQ ID NO;

864), RASQGISSWLA (SEQ ID NO;

865), RASQVSSSYLA (SEQ ID NO;

866), RASQSISSWLA (SEQ ID NO;

867), RSSQSLLDSDDGNTYLD (SEQ ID NO;

868), TGTSSDVGGTNYVS (SEQ ID NO;

869), TLSSDLSVGGKNMF (SEQ ID NO;

870), and an amino acid sequence that exhibits at least about 65% sequence identity to an amino acid sequence of SEQ ID NOS;

861-870;

wherein the antibody or antigen-binding fragment optionally is isolated and optionally further comprises at least one of the following;

g) a CDR that exhibits 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence of amino acids set forth in any of SEQ ID NOS;

830-836 and 844-890; and

/or

h) 2, 3, 4, 5 or 6 different CDRs.

137. An antibody or antigen-binding fragment according to claim 136 that is a full-length antibody, optionally a full-length IgG antibody, or an antibody fragment, optionally an antibody fragment selected from the group consisting of a Fab, Fab′

, F(ab′

)₂, single-chain Fv (scFv), Fv, dsFv, diabody, Fd and Fd′

fragment, Fab fragment, Fd fragment, scFv fragment, and a scFab fragment.

138. An antibody or antigen-binding fragment according to claim 136, wherein a CDR set forth in any of SEQ ID NOS:

830-836 and 844-890 is modified by amino acid replacement, addition, or deletion.

139. An antibody or antigen-binding fragment according to claim 136, wherein:

a) CDRH1 has the amino acid sequence of SEQ ID NO;

830 except that the amino acid residue at position(s) T28, F29, T30, S31, and/or Y33 is different than the corresponding amino acid residue in SEQ ID NO;

830, optionally T28A, F29A, T30A, S31A, and/or Y33A;

or

b) CDRH1 has the amino acid sequence of SEQ ID NO;

831 except that the amino acid residue at position(s) S28, F29, T30, W33, I34, and/or G35 is different than the corresponding amino acid residue in SEQ ID NO;

831, optionally S28A, S28R, S28K, S28N, F29A, T30A, W33A, I34A, G35T, G35A, and/or G35V; and

/or

c) CDRH2 has the amino acid sequence of SEQ ID NO;

844 except that the amino acid residue at position(s) I50, I51, N52, P52a, S53, G54, G55, G56, T57, and/or S58 is different than the corresponding amino acid residue in SEQ ID NO;

844, optionally I50A, I50T, I51A, I51T, I51V, I51N, I51R, I51W, I51S, I51G, I51V, I51E, I51H, I51Y, N52A, N52V, N52G, N52T, N52P, N52L, N52W, N52Y, N52V, N52S, N52Q, N52K, P52aA, P52aM, P52aE, P52aH, P52aY, P52aT, P52aN, P52aR, P52aW, P52aS, P52aG, S53A, S53I, S53E, S53R, S53G, S53T, S53L, S53V, S53N, S53P, G54A, G54W, G54D, G55A, G55V, G55E, G55S, G55K, G55T, G55L, G55R, G55H, G55I, G55W, S58A, T57A, and/or S58A;

or

d) CDRH2 has the amino acid sequence of SEQ ID NO;

844 except for one of the following sets of differences, I51V/N52L/S53T/G55H, N52L/S53T/G55H, I51E/N52L/S53T/G55H, or I51N/N52L/S53T/G55H;

or

e) CDRH2 has the amino acid sequence of SEQ ID NO;

845 except that the amino acid residue at position(s) I50, I51, Y52, P52a, D54, S55, D56 and/or T57 is different than the corresponding amino acid residue in SEQ ID NO;

845, optionally I50A, I51A, Y52A, P52aA, D54A, S55G, D56A, T57D, and/or T57A; and

/or f) CDRH3 has the amino acid sequence of SEQ ID NO;

851 except that the amino acid residue at position(s) E96, Y97, S98, S99, S100, S100a, A100b, E100c, Q101, and/or H102 is different than the corresponding amino acid residue in SEQ ID NO;

851, optionally E96A, Y97A, S98A, S98Q, S98V, S98I, S98G, S99P, S99A, S99L, S99W, S99F, S99N, S99H, S99C, S99G, S100F, S100A, S100G, S100C, S100H, S100L, S100R, S100aA, A100bE, E100cA, Q101A, H102A, H102S, H102F, and/or H102Y;

or

g) CDRH3 has the amino acid sequence of SEQ ID NO;

851 except for one of the following sets of differences, S98A/S99P/S100F, S98A/S99P/S100F/H102F, or S98A/S99P/S100F/H102Y;

or

h) CDRH3 has the amino acid sequence of SEQ ID NO;

852 except that the amino acid residue at position(s) R95, G96, Y97, S98, Y99, G100, Y100a, D100b, and/or D101 is different than the corresponding amino acid residue in SEQ ID NO;

852, optionally R95A, G96K, G96R, G96L, G96D, G96T, Y97A, Y97H, S98A, Y99A, G100A, G100D, G100L, G100P, G100R, G100M, G100K, G100S, G100R, G100T, Y100aA, D100bA, and/or D101A;

or

i) CDRH3 has the amino acid sequence of SEQ ID NO;

852 except for the following differences, G96K/G100T; and

/or

j) CDRL1 has the amino acid sequence of SEQ ID NO;

861 except that the amino acid residue at position(s) R24, Q27, S28, S30, S31, and/or Y32 is different than the corresponding amino acid residue in SEQ ID NO;

861, optionally R24G, Q27L, S28P, S28G, S28K, S28V, S28F, S28P, S28T, S28L, S28Q, S28A, S28N, S28H, S28I, S28R, S28W, S28M, S28E, S30N, S30W, S30R, S30L, S30C, S30D, S30L, S30T, S30P, S30Y, S530Q, S30A, S30G, S30V, S31K, S31T, S31N, S31K, S31L, S31M, S31F, S31I, S31V, S31H, S31A, S31P, S31D, S31R, S31Y, S31Q, S31E, S31G, Y32V, and/or Y32S;

or

k) CDRL1 has the amino acid sequence of SEQ ID NO;

861 except for the following differences, S28N/S30D/S31H;

or

l) CDRL1 has the amino acid sequence of SEQ ID NO;

862 except that the amino acid residue at position(s) G24, L25, S26, S27, G27a, S27b, V27c, S28, T29, S30, Y31, Y32, and/or P33 is different than the corresponding amino acid residue in SEQ ID NO;

862, optionally G24A, G24R, G24L, L25A, S26A, S27A, G27aA, S27bA, V27cA, S28A, T29A, S30A, Y31A, Y32A, and/or P33A; and

/or

m) CDRL2 has the amino acid sequence of SEQ ID NO;

871 except that the amino acid residue at position(s) D50, A51, S52, N53, R54, A55, and/or T56 is different than the corresponding amino acid residue in SEQ ID NO;

871, optionally D50A, A51T, S52A, S52L, S52T, S52R, S52S, S52W, S52N, S52P, S52M, N53A, N53E, N53G, N53M, N53C, N53H, N53P, R54A, A55T, A55R, A55C, A55S, A55G, and/or T56A;

or

n) CDRL2 has the amino acid sequence of SEQ ID NO;

871 except for one of the following sets of differences, S52L/A55S or S52L/A55G;

or

o) CDRL2 has the amino acid sequence of SEQ ID NO;

872 except that the amino acid residue at position(s) S50, T51, N52, T53, R54, S55, and/or S56 is different than the corresponding amino acid residue in SEQ ID NO;

872, optionally S50A, S50F, S50G, S50C, S50R, S50L, S50M, S50V, S50P, S50T, S50H, S50Q, S50N, S50K, S50D, S50E, S50W, T51A, T51F, T51L, T51I, T51M, T51V, T51S, T51P, T51Y, T51H, T51Q, T51N, T51K, T51D, T51E, T51W, T51R, T51G, N52A, T53A, R54A, R54I, R54Y, R54D, R54G, S55A, S55F, S55L, S55I, S55M, S55V, S55P, S55T, S55Y, S55H, S55Q, S55N, S55K, S55D, S55E, S55W, S55R, S55G, and/or S56A; and

/or

p) CDRL3 has the amino acid sequence of SEQ ID NO;

881 except that the amino acid residue at position(s) R91, S92, N93, and/or W94 is different than the corresponding amino acid residue in SEQ ID NO;

881, optionally R91P, R91L, R91G, S92P, S92A, S92Q, S92V, S92T, S92R, S92G, S92V, S92M, S92N, S92C, N93Y, N93S, N93H, N93Q, W94R, W94S, W94T, W94L, W94P, and/or W94M;

or

q) CDRL3 has the amino acid sequence of SEQ ID NO;

882 except that the amino acid residue at position(s) V89, L90, Y91, M92, G93, S94, G95, I95a, and/or S95b is different than the corresponding amino acid residue in SEQ ID NO;

882, optionally V89A, V89P, V89T, V89S, V89L, V89R, V89C, V89E, V89W, V89N, V89I, V89G, V89H, L90A, Y91A, M92A, M92E, M92S, M92G, M92L, M92P, M92V, M92D, M92R, M92N, M92T, M92F, G93A, S94A, S94W, S94G, S94P, S94R, S94L, S94M, S94E, S94V, G94A, I95aA, and/or S95bA;

or

r) CDRL3 has the amino acid sequence of SEQ ID NO;

882 except for one of the following sets of differences, M92R/S94M or V89L/S94P.

140. An antibody or antigen-binding fragment according to claim 136 that comprises:

a) a CDRH2 having the amino acid sequence of SEQ ID NO;

844 except for the difference G55H and a CDRH3 having the amino acid sequence of SEQ ID NO;

except for the differences S98A/S99P/S100F/H102F;

or

b) a CDRH3 having the amino acid sequence of SEQ ID NO;

851 except for the differences S98A/S99P/S100F/H102F and a CDRL1 having the amino acid sequence of SEQ ID NO;

861 except for the differences S28N/S30D/S31H;

or

c) a CDRH2 having the amino acid sequence of SEQ ID NO;

844 except for the differences I51V/N52L/S53T/G55H, a CDRH3 having the amino acid sequence of SEQ ID NO;

851 except for the differences S98A/S99P/S100F/H102F, and a CDRL1 having the amino acid sequence of SEQ ID NO;

861 except for the differences S28N/S30D/S31H;

or

d) a CDRH2 having the amino acid sequence of SEQ ID NO;

844 except for the differences I51V/N52L/S53T/G55H, a CDRH3 having the amino acid sequence of SEQ ID NO;

851 except for the differences S98A/S99P/S100F/H102F, a CDRL1 having the amino acid sequence of SEQ ID NO;

861 except for the differences S28N/S30D/S31H, and a CDRL2 having the amino acid sequence of SEQ ID NO;

871 except for the differences S52L/A55S;

or

e) a CDRH1 having the amino acid sequence of SEQ ID NO;

831 except for the differences S28R/G35V, a CDRH3 having the amino acid sequence of SEQ ID NO;

852 except for the differences G96K/G100T, and a CDRL3 having the amino acid sequence of SEQ ID NO;

882 except for the differences M92R/S94M;

or

f) a CDRH1 having the amino acid sequence of SEQ ID NO;

831 except for the differences S28R/G35V, a CDRH3 having the amino acid sequence of SEQ ID NO;

852 except for the differences G96K/G100T, and a CDRL3 having the amino acid sequence of SEQ ID NO;

882 except for the differences V89L/S94P;

or

g) a CDRH1 having the amino acid sequence of SEQ ID NO;

831 except for the differences S28R/G35V, a CDRH3 having the amino acid sequence of SEQ ID NO;

852 except for the differences G96K/G100T, a CDRL2 having the amino acid sequence of SEQ ID NO;

872 except for the differences S50G, and a CDRL3 having the amino acid sequence of SEQ ID NO;

882 except for the differences V89L/S94P.

141. An antibody or antigen-binding fragment according to claim 136 that further comprises one or more, optionally 2, 3, 4, 5, 6, 7, 8, 9, 10 or more, amino acid differences in a framework region of the antibody or antigen-binding fragment.

142. An antibody or antigen-binding fragment according to claim 141, wherein:

a) each light chain variable domain comprises one or more amino acid differences in a framework region, optionally an amino acid difference at position 62 and/or 76, optionally one or more amino acid differences selected from the group consisting of F62L, S76E, S76Q, S76P, S76L, S76T, S76G, S76A, S76Y, S76N, T76S, T76E, T76Y, and T76M; and

/or

b) each heavy chain variable domain comprises one or more amino acid differences, optionally an amino acid difference at position 24 and/or 82a, optionally one or more amino acid differences selected from the group consisting of G24T, G24L G24A, and S82aT.

143. An antibody or antigen-binding fragment according to claim 136 that exhibits at least a binding affinity that is 10⁻

6M, 10<sup>−

7</sup>M, 10<sup>−

8</sup>M, 10<sup>−

6</sup>M, 10<sup>−

10</sup>M, 10<sup>−

11</sup>M, or 10<sup>−

12</sup>M or lower as measured by surface plasmon resonance (SPR).

144. An antibody or antigen-binding fragment according to claim 136 that is germline-derived, wherein the antibody or antigen-binding fragment optionally comprises:

A. a heavy chain variable domain comprising V_H, D_H, and J_H germline segments or modified germline segments and a light chain variable domain comprising V_κ

and J_κ

or V_λ

and J_λ

germline segments, wherein;

a) the V_H germline segment is an IGHV1, an IGHV4, an IGHV5, or an IGHV6 germline segment or allelic variant thereof, optionally an IGHV1-3*01, an IGHV1-3*02, an IGHV1-8*01, an IGHV1-46*01, an IGHV1-46*02, an IGHV1-46*03, an IGHV4-31*01, an IGHV4-31*02, an IGHV4-31*03, an IGHV4-31*04, an IGHV4-31*05, an IGHV4-31*06, an IGHV4-31*07, an IGHV4-31*08, an IGHV4-31*09, an IGHV4-31*10, an IGHV4-34*01, an IGHV4-34*02, an IGHV4-34*03, an IGHV4-34*04, an IGHV4-34*05, an IGHV4-34*06, an IGHV4-34*07, an IGHV4-34*08, an IGHV4-34*09, an IGHV4-34*10, an IGHV4-34*11, an IGHV4-34*12, an IGHV4-34*13, an IGHV5-51*01, an IGHV5-51*02, an IGHV5-51*03, an IGHV5-51*04, an IGHV5-51*05, an IGHV6-1*01, or an IGHV6-1*02 germline segment or a modification or optimized derivative thereof; and

b) the D_H germline segment is an IGHD6, an IGHD5, an IGHD4, an IGHD2, an IGHD3, or an IGHD7 germline segment or allelic variant thereof, optionally an IGHD2-2*01, an IGHD2-2*02, an IGHD2-15*01, an IGHD4-23*01, an IGHD6-6*01, an IGHD6-13*01, IGHD5-18*01, IGHD3-3*01, an IGHD3-3*02, an IGHD3-10*01, an IGHD3-10*02, or an IGHD7-27*01 germline segment or a modification or optimized derivative thereof; and

c) the J_H germline segment is an IGHJ1, an IGHJ2, an IGHJ4, or an IGHJ6 germline segment or allelic variant thereof, optionally an IGHJ1*01, an IGHJ2*01, an IGHJ4*01, an IGHJ4*02, an IGHJ4*03, an IGHJ6*01, an IGHJ6*02, an IGHJ6*03, or an IGHJ6*04 germline segment or a modification or optimized derivative thereof; and

d) the V_κ

germline segment is an IGKV1, an IGKV2, or an IGKV3 germline segment or allelic variant thereof and the J_κ

is a IGKJ1 germline segment or allelic variant thereof, optionally an IGKV1-5*01, an IGKV1-5*02, an IGKV1-5*03, an IGKV1-12*01, an IGKV1-12*02, an IGKV2-D-40*01, an IGKV3-11*01, an IGKV3-11*02, an IGKV3-15*01, an IGKV3-20*01, or an IGKV3-20*02 germline segment or a modification or optimized derivative thereof and an IGKJ1*01 germline segment or a modification or optimized derivative thereof;

or

e) the V_λ

germline segment is an IGLV2, an IGLV8, an IGLV11, or an IGLV5 germline segment or allelic variant thereof and the J_λ

germline segment is an IGLJ1 or an IGLJ4 germline segment or allelic variant thereof, optionally an IGLV2-14*01, an IGLV2-14*02, an IGLV2-14*03, an IGLV2-14*04, an IGLV8-61*01, an IGLV8-61*02, an IGLV8-61*03, an IGLV5-48*01, or an IGLV11-55*01 germline segment or a modification or optimized derivative thereof and an IGLJ1*01 or IGLJ4*01 germline segment or a modification or optimized derivative thereof; and

/or

B. a heavy chain variable domain comprising V_H, D_H, and J_H germline segments or modified germline segments and a light chain variable domain comprising V_κ

and J_κ

or V_λ

and J_λ

germline segments, wherein;

a) the heavy chain variable domain comprises IGHV1-46*01, an IGHD6-6*01, and an IGHJ1*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV3-11*01 and IGKJ1*01 germline segments or modified forms thereof;

or

b) the heavy chain variable domain comprises IGHV5-51*03, IGHD5-18*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGLV8-61*01 and IGLJ1*01 germline segments or modified forms thereof;

or

c) the heavy chain variable domain comprises IGHV6-1*01, IGHD3-3*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGLV5-48*01 and IGLJ4*01 germline segments or a modified forms thereof;

or

d) the heavy chain variable domain comprises IGHV1-46*01, IGHD6-13*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV3-15*01 and IGKJ1*01 germline segments or modified forms thereof;

or

e) the heavy chain variable domain comprises IGHV4-34*01, IGHD7-27*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV1-12*01 and IGKJ1*01 germline segments or modified forms thereof;

or

f) the heavy chain variable domain comprises IGHV1-46*01, IGHD6-13*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV3-20*01 and IGKJ1*01 germline segments or modified forms thereof;

or

g) the heavy chain variable domain comprises IGHV1-3*02, IGHD4-23*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV1-5*01 and IGKJ1*01 germline segments or modified forms thereof;

or

h) the heavy chain variable domain comprises IGHV1-46*01, IGHD2-15*01, and IGHJ2*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV1-5*01 and IGKJ1*01 germline segments or modified forms thereof;

or

i) the heavy chain variable domain comprises IGHV1-46*01, IGHD3-10*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV1-5*01 and IGKJ1*01 germline segments or modified forms thereof;

or

j) the heavy chain variable domain comprises IGHV1-8*01, IGHD2-2*01, and IGHJ6*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV1-5*01 and IGKJ1*01 germline segments or modified forms thereof;

or

k) the heavy chain variable domain comprises IGHV1-46*01, IGHD6-13*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGKV2D-40*01 and, IGKJ1*01 germline segments or modified forms thereof;

or

l) the heavy chain variable domain comprises IGHV4-34*01, IGHD7-27*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGLV2-14*01 and IGLJ4*01 germline segments or modified forms thereof;

or

m) the heavy chain variable domain comprises IGHV4-31*02, IGHD2-15*01, and IGHJ2*01 germline segments or modified forms thereof and the light chain variable domain comprises IGLV2-14*01 and IGLJ4*01 germline segments or modified forms thereof;

or

n) the heavy chain variable domain comprises IGHV4-34*01, IGHD7-27*01, and IGHJ4*01 germline segments or modified forms thereof and the light chain variable domain comprises IGLV11-55*01 and IGLJ4*01 germline segments or a modified form thereof; and

/or

C. a heavy chain variable domain comprising V_H, D_H, and J_H germline segments or modified germline segments and a light chain variable domain comprising V_κ

and J_κ

or V_λ

and J_λ

germline segments, wherein;

a) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

131 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

b) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

132 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

142;

or

c) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

133 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

143;

or

d) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

135 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

145;

or

e) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

137 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

146;

or

f) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

135 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

144;

or

g) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

138 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

147;

or

h) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

136 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

147;

or

i) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

134 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

147;

or

j) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

139 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

147;

or

k) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

135 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

148;

or

l) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

137 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

149;

or

m) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

140 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

149;

or

n) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

137 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

150.

D. a heavy chain variable domain comprising V_H, D_H, and J_H germline segments or modified germline segments and a light chain variable domain comprising V_κ

and J_κ

or V_λ

and J_λ

germline segments, wherein;

a) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

155 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

b) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

156 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

c) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

385 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

142.

E. a heavy chain variable domain comprising V_H, D_H, and J_H germline segments or modified germline segments and a light chain variable domain comprising V_κ

and J_κ

or V_λ

and J_λ

germline segments, wherein;

a) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

384 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

142;

or

b) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

414 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

142;

or

c) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

433 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

142;

or

d) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

433 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

479;

or

e) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

433 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

537;

or

f) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

433 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

536;

or

g) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

131 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

h) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

151 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

i) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

155 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

j) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

156 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

k) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

157 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

l) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

155 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

266;

or

m) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

219 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

141;

or

n) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

156 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

343;

or

o) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

239 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

343;

or

p) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

239 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

370;

or

q) the heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

134 and the light chain variable domain comprises an amino acid sequence of SEQ ID NO;

147;

or

F. a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO;

433 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO;

537;

or

G. a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO;

433 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO;

536;

or

H. a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO;

239 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO;

343;

or

I. a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO;

239 and a light chain variable domain comprising an amino acid sequence of SEQ ID NO;

370;

or

145. An antibody or antigen-binding fragment according to claim 136, wherein the antibody or antigen-binding fragment (i) exhibits affinity for human DLL4 expressed on the surface of a cell, optionally on the surface of an endothelial cell, and/or (ii) modulates an activity of human DLL4, optionally as an agonist or antagonist, wherein such modulation optionally comprises inhibiting Notch activation.

146. An antibody or antigen-binding fragment according to claim 136, wherein the antibody or antigen-binding fragment specifically binds to an epitope within an EGF domain of DLL4, optionally within the EGF2 to EGF4 domains of DLL4, optionally within the EGF3 to EGF4 domain or in EGF2 domain of DLL4.

147. An antibody or antigen-binding fragment according to claim 136, wherein the antibody or antigen-binding fragment specifically binds to a human DLL4 epitope within amino acids 283 to 360 of human DLL4 set forth in SEQ ID NO:

114, wherein the antibody or antigen-binding fragment comprises;

A. at least one heavy chain variable domain and at least one light chain variable domain, wherein;

a) each heavy chain variable domain comprises (i) a CDRH1 having an amino acid sequence GYSFTSYWIG (SEQ ID NO;

831) or an amino acid sequence that has at least 65% sequence identity to SEQ ID NO;

831, (ii) a CDRH2 having an amino acid sequence IIYPGDSDTRYSPSFQG (SEQ ID NO;

845) or an amino acid sequence that has at least 65% sequence identity to SEQ ID NO;

845; and

(iii) a CDRH3 having an amino acid sequence RGYSYGYDYFDY(SEQ ID NO;

852) or an amino acid sequence that has at least 65% sequence identity to SEQ ID NO;

852; and

b) each light chain variable domain comprises (i) a CDRL1 having an amino acid sequence GLSSGSVSTSYYPS(SEQ ID NO;

862) or an amino acid sequence that has at least 65% sequence identity to SEQ ID NO;

862, (ii) a CDRL2 having an amino acid sequence STNTRSS (SEQ ID NO;

872) or an amino acid sequence that has at least 65% sequence identity to SEQ ID NO;

872, and (iii) a CDRL3 having an amino acid sequence VLYMGSGISYV (SEQ ID NO;

882) or an amino acid sequence that has at least 65% sequence identity to SEQ ID NO;

882,

wherein the antibody or antigen-binding antibody fragment is optionally a full-length antibody, optionally a full-length IgG antibody, or an antibody fragment selected from the group consisting of a Fab, Fab′

, F(ab′

)₂, single-chain Fv (scFv), Fv, dsFv, diabody, Fd and Fd′

fragment, Fab fragment, Fd fragment, scFv fragment, and a scFab fragment;

or

B. at least one at least one heavy chain variable domain and at least one light chain variable domain, wherein each heavy chain variable domain comprises three complementary determining regions (CDRs), CDRH1, CDRH2, and CDRH3, and each light chain variable domain comprises three CDRs, CDRL1, CDRL2, and CDRL3, wherein;

a) CDRH1 comprises a modification of amino acid sequence SEQ ID NO;

831, wherein one or more amino acids are different at positions S28, F29, T30, W33, I34, and/or G35 as compared to the corresponding positions in SEQ ID NO;

831, wherein optionally one or more of the differences is selected from the group consisting of S28A, S28R, S28K, S28N, F29A, T30A, W33A, I34A, G35T, G35A and G35V;

b) CDRH2 comprises a modification of amino acid sequence SEQ ID NO;

845, wherein one or more amino acids are different at positions I50, I51, Y52, P52a, D54, S55, D56, and/or T57 as compared to the corresponding positions in SEQ ID NO;

845, wherein optionally one or more of the differences is selected from the group consisting of I50A, I51A, Y52A, P52aA, D54A, S55G, D56A, T57D, and T57A;

c) CDRH3 comprises a modification of amino acid sequence SEQ ID NO;

852, wherein one or more amino acids are different at positions R95, G96, Y97, S98, Y99, G100, Y100a, D100b, and/or D101 as compared to the corresponding positions in SEQ ID NO;

852, wherein optionally one or more of the differences is selected from the group consisting of R95A, G96K, G96R, G96L, G96D, G96T, Y97A, Y97H, S98A, Y99A, G100A, G100D, G100L, G100P, G100R, G100M, G100K, G100S, G100R, G100T, Y100aA, D100bA and D101A, and wherein optionally the differences are G96K/G100T;

d) CDRL1 comprises a modification of amino acid sequence SEQ ID NO;

862, wherein one or more amino acids are different at positions G24, L25, S26, S27, G27a, S27b, V27c, S28, T29, S30, Y31, Y32, and/or P33 as compared to the corresponding positions in SEQ ID NO;

862, wherein optionally one or more of the differences is selected from the group consisting of G24A, G24R, G24L, L25A, S26A, S27A, G27aA, S27bA, V27cA, S28A, T29A, S30A, Y31A, Y32A, and P33A;

e) CDRL2 comprises a modification of amino acid sequence SEQ ID NO;

872, wherein one or more amino acids are different at positions S50, T51, N52, T53, R54, S55 and S56 as compared to the corresponding positions in SEQ ID NO;

872, wherein optionally one or more of the differences is selected from the group consisting of S50A, S50F, S50G, S50C, S50R, S50L, S50M, S50V, S50P, S50T, S50H, S50Q, S50N, S50K, S50D, S50E, S50W, T51A, T51F, T51L, T51I, T51M, T51V, T51S, T51P, T51Y, T51H, T51Q, T51N, T51K, T51D, T51E, T51W, T51R, T51G, N52A, T53A, R54A, R54I, R54Y, R54D, R54G, S55A, S55F, S55L, S55I, S55M, S55V, S55P, S55T, S55Y, S55H, S55Q, S55N, S55K, S55D, S55E, S55W, S55R, S55G, and S56A; and

f) CDRL3 comprises a modification of amino acid sequence SEQ ID NO;

882, wherein one or more amino acids are different at positions V89, L90, Y91, M92, G93, S94, G95, I95a, and/or S95b as compared to the corresponding positions in SEQ ID NO;

882, wherein optionally one or more of the differences is selected from the group consisting of V89A, V89P, V89T, V89S, V89L, V89R, V89C, V89E, V89W, V89N, V89I, V89G, V89H, L90A, Y91A, M92A, M92E, M92S, M92G, M92L, M92P, M92V, M92D, M92R, M92N, M92T, M92F, G93A, S94A, S94W, S94G, S94P, S94R, S94L, S94M, S94E, S94V, G94A, I95aA, and S95bA, and wherein optionally the differences are M92R/S94M or V89L/S94P;

or

C. at least one at least one heavy chain variable domain and at least one light chain variable domain, wherein each heavy chain variable domain comprises an amino acid sequence of SEQ ID NO;

433 and each light chain variable domain comprises an amino acid sequence of SEQ ID NO;

537.

148. A composition comprising a first polynucleotide that comprises a coding sequence for a heavy chain variable domain of an antibody or antigen-binding fragment and a second polynucleotide that comprises a coding sequence for a light chain variable domain of the antibody or antigen-binding fragment, which is an antibody or antigen-binding fragment according to claim 136, and wherein the composition optionally comprises at least one of the following:

a) a first vector that contains the first polynucleotide, wherein the first vector optionally is an expression vector;

b) a second vector that contains the second polynucleotide, wherein the second vector optionally is an expression vector;

c) a vector that contains the first and second polynucleotides, wherein the vector optionally is an expression vector;

d) a host cell, optionally a prokaryotic or eukaryotic host cell, stably transfected with the first polynucleotide, wherein the host cell optionally expresses the first polynucleotide to produce a polypeptide that comprises the heavy chain variable domain of an antibody or antigen-binding fragment encoded by the first polynucleotide;

e) a host cell, optionally a prokaryotic or eukaryotic host cell, stably transfected with the second polynucleotide, wherein the host cell optionally expresses the second polynucleotide to produce a polypeptide that comprises the light chain variable domain of an antibody or antigen-binding fragment encoded by the second polynucleotide; and

f) a host cell, optionally a prokaryotic or eukaryotic host cell, stably transfected with the first and second polynucleotides, wherein the host cell optionally expresses the first and second polynucleotides to produce a first polypeptide that comprises the heavy chain variable domain of an antibody or antigen-binding fragment encoded by the first polynucleotide and a second polypeptide that comprises the light chain variable domain of an antibody or antigen-binding fragment encoded by the second polynucleotide, and wherein the first and second polypeptides optionally assemble to form an antibody or antigen-binding fragment.

149. A method of making an anti-DLL4 antibody or antigen-binding antibody fragment, comprising expressing in a suitable host cell a composition according to claim 148 and recovering the antibody or antigen-binding antibody fragment.

150. A composition comprising a carrier, optionally a pharmaceutically acceptable carrier, and an isolated antibody or antigen-binding antibody fragment according to claim 136, wherein the composition is optionally formulated into a formulation for single dose administration or as a sustained release formulation, wherein the formulation is a gel, ointment, cream, paste, suppository, flush, liquid, suspension, aerosol, tablet, pill, or powder, which formulation is optionally suitable for systemic, parenteral, topical, oral, mucosal, intranasal, subcutaneous, aerosolized, intravenous, bronchial, pulmonary, vaginal, vulvovaginal, or esophageal administration,

and wherein the composition is optionally further combined with anti-angiogenic agent, optionally an antagonist of vascular endothelial growth factor (VEGF), wherein the VEGF antagonist optionally is an anti-VEGF antibody or antigen-binding antibody fragment, wherein the anti-VEGF antibody optionally is bevacizumab.

151. A method of treatment, comprising administering a therapeutically effective amount of an antibody or antigen-binding antibody fragment according to claim 136 to a subject in need of treatment for a disease or disorder associated with expression and/or activity of DLL4, wherein the disease or disorder is selected from the group consisting of a tumor;

cancer, optionally prostate cancer, pancreatic cancer, colon cancer, lung cancer, or breast cancer;

atherosclerosis;

arthritis;

ocular neovascularization;

endometriosis;

uterine fibroids;

pre-eclampsia;

or a cell proliferative disorder,

and wherein the method optionally further comprises at least one of the following;

a) administering an effective amount of an anti-angiogenic agent, optionally an antagonist of vascular endothelial growth factor (VEGF), wherein the VEGF antagonist optionally is an anti-VEGF antibody or antigen-binding antibody fragment, wherein the anti-VEGF antibody optionally is bevacizumab; and

/or

b) administering an effective amount of a chemotherapeutic agent.

152. A method of inhibiting Notch activation, comprising delivering an antibody or antigen-binding antibody fragment according to claim 136 in an amount that inhibits Notch activation.