Dual variable domain immunoglobulin and uses thereof

• US 20070071675A1
• Filed: 08/18/2006
• Published: 03/29/2007
• Est. Priority Date: 08/19/2005
• Status: Active Grant

• Current Assignee: Abbvie Incorporated
• Original Assignee: Abbott Laboratories Incorporated
• Inventors: Wu, Chengbin, Salfeld, Jochen, Ghayur, Tariq, Dixon, Richard
• Granted Patent: US 7,612,181 B2
• Time in Patent Office: Days
• US Class Current: 424/1.490
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/42 Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein oligopeptides having up to five amino acids A61K47/183; polyamino acids A61K47/34
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61K 51/1093 conjugates with carriers being antibodies
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 23/00 Anaesthetics
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/06 : Antimigraine agents
• CPC Class Codes: A61P 25/16 : Anti-Parkinson drugs
• CPC Class Codes: A61P 25/18 : Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/24 : Antidepressants
• CPC Class Codes: A61P 25/28 : for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 25/32 : Alcohol-abuse
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 41/00 : Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• 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 16/22 : against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/24 : against cytokines, lymphokines or interferons
• CPC Class Codes: C07K 16/241 : Tumor Necrosis Factors
• CPC Class Codes: C07K 16/244 : Interleukins [IL]
• CPC Class Codes: C07K 16/245 : IL-1
• CPC Class Codes: C07K 16/2809 : against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2887 : against CD20
• CPC Class Codes: C07K 16/2896 : against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/40 : against enzymes
• CPC Class Codes: C07K 16/46 : Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• CPC Class Codes: C07K 16/467 : Igs with modifications in the FR-residues only
• CPC Class Codes: C07K 16/468 : Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 : multispecific
• CPC Class Codes: C07K 2317/51 : Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/522 : CH1 domain
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/64 : comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention relates to engineered multivalent and multispecific binding proteins, methods of making, and specifically to their uses in the prevention and/or treatment of acute and chronic inflammatory and other diseases.

Claims

1. A binding protein comprising a polypeptide chain, wherein said polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first variable domain, VD2 is a second variable domain, C is a constant domain, X1 represents an amino acid or polypeptide, X2 represents an Fc region and n is 0 or 1.

2. The binding protein according to claim 1, wherein said VD1 and VD2 are heavy chain variable domains.

3. The binding protein according to claim 2, wherein said heavy chain variable domain is selected from the group consisting of a murine heavy chain variable domain, a human heavy chain variable domain, a CDR grafted heavy chain variable domain, and a humanized heavy chain variable domain.

4. The binding protein according to claim 2, wherein VD1 and VD2 are capable of binding the same antigen.

5. The binding protein according to claim 2, wherein VD1 and VD2 are capable of binding different antigens.

6. The binding protein according to claim 2, wherein C is a heavy chain constant domain.

7. The binding protein according to claim 6, wherein X1 is a linker with the proviso that X1 is not CH1.

8. The binding protein according to claim 7, wherein the linker is selected from the group consisting of AKTTPKLEEGEFSEAR;

AKTTPKLEEGEFSEARV;

AKTTPKLGG;

SAKTTPKLGG;

AKTTPKLEEGEFSEARV;

SAKTTP;

SAKTTPKLGG;

RADAAP;

RADAAPTVS;

RADAAAAGGPGS;

RADAAAA(G₄S)₄;

SAKTTP;

SAKTTPKLGG;

SAKTTPKLEEGEFSEARV;

ADAAP;

ADAAPTVSIFPP;

TVAAP;

TVAAPSVFIFPP;

QPKAAP;

QPKAAPSVTLFPP;

AKTTPP;

AKTTPPSVTPLAP;

AKTTAP;

AKTTAPSVYPLAP;

ASTKGP;

ASTKGPSVFPLAP;

GGGGSGGGGSGGGGS;

GENKVEYAPALMALS;

GPAKELTPLKEAKVS; and

GHEAAAVMQVQYPAS.

9. The binding protein according to claim 7, wherein X2 is an Fc region.

10. The binding protein according to claim 9, wherein said Fc region is a variant Fc region.

12. The binding protein according to claim 1, wherein said VD1 and VD2 are light chain variable domains.

13. The binding protein according to claim 12, wherein said light chain variable domain is selected from the group consisting of a murine light chain variable domain, a human light chain variable domain, a CDR grafted light chain variable domain, and a humanized light chain variable domain.

14. The binding protein according to claim 12, wherein VD1 and VD2 are capable of binding the same antigen.

15. The binding protein according to claim 12, wherein VD1 and VD2 are capable of binding different antigens.

16. The binding protein according to claim 12, wherein C is a light chain constant domain.

17. The binding protein according to claim 16, wherein X1 is a linker with the proviso that X1 is not CL1.

18. The binding protein according to claim 17, wherein the linker is selected from the group consisting of AKTTPKLEEGEFSEAR;

AKTTPKLEEGEFSEARV;

AKTTPKLGG;

SAKTTPKLGG;

AKTTPKLEEGEFSEARV;

SAKTTP;

SAKTTPKLGG;

RADAAP;

RADAAPTVS;

RADAAAAGGPGS;

RADAAAA(G₄S)₄, SAKTTP;

SAKTTPKLGG;

SAKTTPKLEEGEFSEARV;

ADAAP;

ADAAPTVSIFPP;

TVAAP;

TVAAPSVFIFPP;

QPKAAP;

QPKAAPSVTLFPP;

AKTTPP;

AKTTPPSVTPLAP;

AKTTAP;

AKTTAPSVYPLAP;

ASTKGP;

ASTKGPSVFPLAP;

GGGGSGGGGSGGGGS;

GENKVEYAPALMALS;

GPAKELTPLKEAKVS; and

GHEAAAVMQVQYPAS.

19. The binding protein according to claim 17, wherein the binding protein does not comprise X2.

23. The binding protein according to claim 1, wherein said binding protein is capable of binding one or more targets.

24. The binding protein according to claim 23, wherein the target is selected from the group consisting of ABCF1;

ACVR1;

ACVR1B;

ACVR2;

ACVR2B;

ACVRL1;

ADORA2A;

Aggrecan;

AGR2;

AICDA;

AIF1;

AIG1;

AKAP1;

AKAP2;

AMH;

AMHR2;

ANGPT1;

ANGPT2;

ANGPTL3;

ANGPTL4;

ANPEP;

APC;

APOC1;

AR;

AZGP1 (zinc-a-glycoprotein);

B7.1;

B7.2;

BAD;

BAFF;

BAG1;

BAI1;

BCL2;

BCL6;

BDNF;

BLNK;

BLR1 (MDR15);

BlyS;

BMP1;

BMP2;

BMP3B (GDF10);

BMP4;

BMP6;

BMP8;

BMPR1A;

BMPR1B;

BMPR2;

BPAG1 (plectin);

BRCA1;

C19orf10 ( IL27w);

C3;

C4A;

C5;

C5R1;

CANT1;

CASP1;

CASP4;

CAV1;

CCBP2 (D6/JAB61);

CCL1 (I-309);

CCL11 (eotaxin);

CCL13 (MCP-4);

CCL15 (MIP-1d);

CCL16 (HCC-4);

CCL17 (TARC);

CCL18 (PARC);

CCL19 (MIP-3b);

CCL2 (MCP-1);

MCAF;

CCL20 (MIP-3a);

CCL21 (MIP-2);

SLC;

exodus-2;

CCL22 (MDC/STC-1);

CCL23 (MPIF-1);

CCL24 (MPIF-2/eotaxin-2);

CCL25 (TECK);

CCL26 (eotaxin-3);

CCL27 (CTACK/ILC);

CCL28;

CCL3 (MIP-1a);

CCL4 (MIP-1b);

CCL5 (RANTES);

CCL7 (MCP-3);

CCL8 (mcp-2);

CCNA1;

CCNA2;

CCND1;

CCNE1;

CCNE2;

CCR1 (CKR1/HM145);

CCR2 (mcp-1RB/RA);

CCR3 (CKR3/CMKBR3);

CCR4;

CCR5 (CMKBR5/ChemR13);

CCR6 (CMKBR6/CKR-L3/STRL22/DRY6);

CCR7 (CKR7/EBI1);

CCR8 (CMKBR8/TER1/CKR-L1);

CCR9 (GPR-9-6);

CCRL1 (VSHK1);

CCRL2 (L-CCR);

CD164;

CD19;

CD1C;

CD20;

CD200;

CD-22;

CD24;

CD28;

CD3;

CD37;

CD38;

CD3E;

CD3G;

CD3Z;

CD4;

CD40;

CD40L;

CD44;

CD45RB;

CD52;

CD69;

CD72;

CD74;

CD79A;

CD79B;

CD8;

CD80;

CD81;

CD83;

CD86;

CDH1 (E-cadherin);

CDH10;

CDH12;

CDH13;

CDH18;

CDH19;

CDH20;

CDH5;

CDH7;

CDH8;

CDH9;

CDK2;

CDK3;

CDK4;

CDK5;

CDK6;

CDK7;

CDK9;

CDKN1A (p21Wap1/Cip1);

CDKN1B (p27Kip1);

CDKN1C;

CDKN2A (p16INK4a);

CDKN2B;

CDKN2C;

CDKN3;

CEBPB;

CER1;

CHGA;

CHGB;

Chitinase;

CHST10;

CKLFSF2;

CKLFSF3;

CKLFSF4;

CKLFSF5;

CKLFSF6;

CKLFSF7;

CKLFSF8;

CLDN3;

CLDN7 (claudin-7);

CLN3;

CLU (clusterin);

CMKLR1;

CMKOR1 (RDC1);

CNR1;

COL18A1;

COL1A1;

COL4A3;

COL6A1;

CR2;

CRP;

CSF1(M-CSF);

CSF2 (GM-CSF);

CSF3 (GCSF);

CTLA4;

CTNNB1 (b-catenin);

CTSB (cathepsin B);

CX3CL1 (SCYD1);

CX3CR1 (V28);

CXCL1 (GRO1);

CXCL10(IP-10);

CXCL11(I-TAC/IP-9);

CXCL12 (SDF1);

CXCL13;

CXCL14;

CXCL16;

CXCL2 (GRO2);

CXCL3 (GRO3);

CXCL5 (ENA-78/LIX);

CXCL6 (GCP-2);

CXCL9 (MIG);

CXCR3 (GPR9/CKR-L2);

CXCR4;

CXCR6 (TYMSTR/STRL33/Bonzo);

CYB5;

CYC1;

CYSLTR1;

DAB21P;

DES;

DKFZp451J0118;

DNCL1;

DPP4;

E2F1;

ECGF1;

EDG1;

EFNA1;

EFNA3;

EFNB2;

EGF;

EGFR;

ELAC2;

ENG;

ENO1;

ENO2;

ENO3;

EPHB4;

EPO;

ERBB2 (Her-2);

EREG;

ERK8;

ESR1;

ESR2;

F3 (TF);

FADD;

FasL;

FASN;

FCER1A;

FCER2;

FCGR3A;

FGF;

FGF1 (aFGF);

FGF10;

FGF11;

FGF12;

FGF12B;

FGF13;

FGF14;

FGF16;

FGF17;

FGF18;

FGF19;

FGF2 (bFGF);

FGF20;

FGF21;

FGF22;

FGF23;

FGF3 (int-2);

FGF4 (HST);

FGF5;

FGF6 (HST-2);

FGF7 (KGF);

FGF8;

FGF9;

FGFR3;

FIGF (VEGFD);

FIL1 (EPSILON);

FIL1 (ZETA);

FLJ12584;

FLJ25530;

FLRT1 (fibronectin);

FLT1;

FOS;

FOSL1 (FRA-1);

FY (DARC);

GABRP (GABAa);

GAGEB1;

GAGEC1;

GALNAC4S-6ST;

GATA3;

GDF5;

GFI1;

GGT1;

GM-CSF;

GNAS1;

GNRH1;

GPR2 (CCR10);

GPR31;

GPR44;

GPR81 (FKSG80);

GRCC10 (C10);

GRP;

GSN (Gelsolin);

GSTP1;

HAVCR2;

HDAC4;

HDAC5;

HDAC7A;

HDAC9;

HGF;

HIF1A;

HIP1;

histamine and histamine receptors;

HLA-A;

HLA-DRA;

HM74;

HMOX1;

HUMCYT2A;

ICEBERG;

ICOSL;

ID2;

IFN-a;

IFNA1;

IFNA2;

IFNA4;

IFNA5;

IFNA6;

IFNA7;

IFNB1;

IFNgamma;

IFNW1;

IGBP1;

IGF1;

IGF1R;

IGF2;

IGFBP2;

IGFBP3;

IGFBP6;

IL-1;

IL10;

IL10RA;

IL10RB;

IL11;

IL11RA;

IL-12;

IL12A;

IL12B;

IL12RB1;

IL12RB2;

IL13;

IL13RA1;

IL13RA2;

IL14;

IL15;

IL15RA;

IL16;

IL17;

IL17B;

IL17C;

IL17R;

IL18;

IL18BP;

IL18R1;

IL18RAP;

IL19;

IL1A;

IL1B;

IL1F10;

IL1F5;

IL1F6;

IL1F7;

IL1F8;

IL1F9;

IL1HY1;

IL1R1;

IL1R2;

IL1RAP;

IL1RAPL1;

IL1RAPL2;

IL1RL1;

IL1RL2 IL1RN;

IL2;

IL20;

IL20RA;

IL21R;

IL22;

IL22R;

IL22RA2;

IL23;

IL24;

IL25;

IL26;

IL27;

IL28A;

IL28B;

IL29;

IL2RA;

IL2RB;

IL2RG;

IL3;

IL30;

IL3RA;

IL4;

IL4R;

IL5;

IL5RA;

IL6;

IL6R;

IL6ST (glycoprotein

     130);

IL7;

IL7R;

IL8;

IL8RA;

IL8RB;

IL8RB;

IL9;

IL9R;

ILK;

INHA;

INHBA;

INSL3;

INSL4;

IRAK1;

IRAK2;

ITGA1;

ITGA2;

ITGA3;

ITGA6 (a6 integrin);

ITGAV;

ITGB3;

ITGB4 (b 4 integrin);

JAG1;

JAK1;

JAK3;

JUN;

K6HF;

KAI1;

KDR;

KITLG;

KLF5 (GC Box BP);

KLF6;

KLK10;

KLK12;

KLK13;

KLK14;

KLK15;

KLK3;

KLK4;

KLK5;

KLK6;

KLK9;

KRT1;

KRT19 (Keratin

     19);

KRT2A;

KRTHB6 (hair-specific type II keratin);

LAMA5;

LEP (leptin);

Lingo-p75;

Lingo-Troy;

LPS;

LTA (TNF-b);

LTB;

LTB4R (GPR16);

LTB4R2;

LTBR;

MACMARCKS;

MAG or Omgp;

MAP2K7 (c-Jun);

MDK;

MIB1;

midkine;

MIF;

MIP-2;

MKI67 (Ki-67);

MMP2;

MMP9;

MS4A1;

MSMB;

MT3 (metallothionectin-III);

MTSS1;

MUC1 (mucin);

MYC;

MYD88;

NCK2;

neurocan;

NFKB1;

NFKB2;

NGFB (NGF);

NGFR;

NgR-Lingo;

NgR-Nogo66 (Nogo);

NgR-p75;

NgR-Troy;

NME1 (NM23A);

NOX5;

NPPB;

NR0B1;

NR0B2;

NR1D1;

NR1D2;

NR1H2;

NR1H3;

NR1H4;

NR1I2;

NR113;

NR2C1;

NR2C2;

NR2E1;

NR2E3;

NR2F1;

NR2F2;

NR2F6;

NR3C1;

NR3C2;

NR4A1;

NR4A2;

NR4A3;

NR5A1;

NR5A2;

NR6A1;

NRP1;

NRP2;

NT5E;

NTN4;

ODZ1;

OPRD1;

P2RX7;

PAP;

PART1;

PATE;

PAWR;

PCA3;

PCNA;

PDGFA;

PDGFB;

PECAM1;

PF4 (CXCL4);

PGF;

PGR;

phosphacan;

PIAS2;

PIK3CG;

PLAU (uPA);

PLG;

PLXDC1;

PPBP (CXCL7);

PPID;

PR1;

PRKCQ;

PRKD1;

PRL;

PROC;

PROK2;

PSAP;

PSCA;

PTAFR;

PTEN;

PTGS2 (COX-2);

PTN;

RAC2 (p21Rac2);

RARB;

RGS1;

RGS13;

RGS3;

RNF110 (ZNF144);

ROBO2;

S100A2;

SCGB1D2 (lipophilin B);

SCGB2A1 (mammaglobin

     2);

SCGB2A2 (mammaglobin

     1);

SCYE1 (endothelial Monocyte-activating cytokine);

SDF2;

SERPINA1;

SERPINA3;

SERPINB5 (maspin);

SERPINE1 (PAI-1);

SERPINF1;

SHBG;

SLA2;

SLC2A2;

SLC33A1;

SLC43A1;

SLIT2;

SPP1;

SPRR1B (Spr1);

ST6GAL1;

STAB1;

STAT6;

STEAP;

STEAP2;

TB4R2;

TBX21;

TCP10;

TDGF1;

TEK;

TGFA;

TGFB11;

TGFB1I1;

TGFB2;

TGFB3;

TGFB1;

TGFBR1;

TGFBR2;

TGFBR3;

TH1L;

THBS1 (thrombospondin-1);

THBS2;

THBS4;

THPO;

TIE (Tie-1);

TIMP3;

tissue factor;

TLR10;

TLR2;

TLR3;

TLR4;

TLR5;

TLR6;

TLR7;

TLR8;

TLR9;

TNF;

TNF-a;

TNFAIP2 (B94);

TNFAIP3;

TNFRSF1A;

TNFRSF1A;

TNFRSF1B;

TNFRSF21;

TNFRSF5;

TNFRSF6 (Fas);

TNFRSF7;

TNFRSF8;

TNFRSF9;

TNFSF10 (TRAIL);

TNFSF11 (TRANCE);

TNFSF12 (APO3L);

TNFSF13 (April);

TNFSF13B;

TNFSF14 (HVEM-L);

TNFSF15 (VEGI);

TNFSF18;

TNFSF4 (OX40 ligand);

TNFSF5 (CD40 ligand);

TNFSF6 (FasL);

TNFSF7 (CD27 ligand);

TNFSF8 (CD30 ligand);

TNFSF9 (4-1BB ligand);

TOLLIP;

Toll-like receptors;

TOP2A (topoisomerase Iia);

TP53;

TPM1;

TPM2;

TRADD;

TRAF1;

TRAF2;

TRAF3;

TRAF4;

TRAF5;

TRAF6;

TREM1;

TREM2;

TRPC6;

TSLP;

TWEAK;

VEGF;

VEGFB;

VEGFC;

versican;

VHL C5;

VLA-4;

XCL1 (lymphotactin);

XCL2 (SCM-1b);

XCR1 (GPR5/CCXCR1);

YY1; and

ZFPM2.

25. The binding protein according to claim 1, wherein said binding protein is capable of binding a two targets, wherein the two targets are selected from the group consisting of CD138 and CD20;

CD138 and CD40;

CD20 and CD3;

CD38 &

CD138;

CD38 and CD20;

CD38 and CD40;

CD40 and CD20;

CD19 and CD20;

CD-8 and IL-6;

PDL-1 and CTLA-4;

CTLA-4 and BTNO2;

CSPGs and RGM A;

IGF1 and IGF2;

IGF1/2 and Erb2B;

IL-12 and IL-18;

IL-12 and TWEAK;

IL-13 and ADAM8;

IL-13 and CL25;

IL-13 and IL-1beta;

IL-13 and IL-25;

IL-13 and IL-4;

IL-13 and IL-5;

IL-13 and IL-9;

IL-13 and LHR agonist;

IL-13 and MDC;

IL-13 and MIF;

IL-13 and PED2;

IL-13 and SPRR2a;

IL-13 and SPRR2b;

IL-13 and TARC;

IL-13 and TGF-β

;

IL-1α and

IL-1β

;

MAG and RGM A;

NgR and RGM A;

NogoA and RGM A;

OMGp and RGM A;

RGM A and RGM B;

Te38 and TNFα

;

TNFα and

IL-12;

TNFα and

IL-12p40;

TNFα and

IL-13;

TNFα and

IL-15;

TNFα and

IL-17;

TNFα and

IL-18;

TNFα and

IL-1beta;

TNFα and

IL-23;

TNFα and

MIF;

TNFα and

PEG2;

TNFα and

PGE4;

TNFα and

VEGF; and

VEGFR and EGFR;

TNFα and

RANK ligand;

TNFα and

Blys;

TNFα and

GP130;

TNFα and

CD-22; and

TNFα and

CTLA-4.

26. The binding protein according to claim 23, wherein the binding protein is capable of modulating a biological function of one or more targets.

27. The binding protein according to claim 23, wherein the binding protein is capable of neutralizing one or more targets.

28. The binding protein according to any one of claims 23-25, wherein the target is selected from the group consisting of cytokine, chemokine, cell surface protein, enzyme and receptor.

29. The binding protein according to claim 28 wherein the cytokine is selected from the group consisting of lymphokines, monokines, and polypeptide hormones.

30. The binding protein according to claim 29, wherein said cytokines are IL-1α

and IL-1β

.

31. The binding protein according to claim 30, wherein the binding protein comprises a DVD heavy chain amino acid sequence selected from the group consisting of SEQ ID NO. 33, SEQ ID NO. 37, SEQ ID NO. 41, SEQ ID NO. 45, SEQ ID NO. 47, SEQ ID NO. 51, SEQ ID NO. 53, SEQ ID NO. 55, SEQ ID NO. 57, and SEQ ID NO. 59;

and a DVD light chain amino acid sequence selected from the group consisting of SEQ ID NO. 35, SEQ ID NO. 39, SEQ ID NO. 43, SEQ ID NO. 46, SEQ ID NO. 49, SEQ ID NO. 52, SEQ ID NO. 54, SEQ ID NO. 56, SEQ ID NO. 58, and SEQ ID NO. 60.

32. The binding protein according to claim 29, wherein said cytokines are TNF-α

and IL-13.

33. The binding protein according to claim 29, wherein said cytokines are IL-12 and IL-18.

34. The binding protein according to claim 33, wherein the binding protein comprises a DVD heavy chain amino acid sequence selected from the group consisting of SEQ ID NO. 83, SEQ ID NO. 90, SEQ ID NO. 93, SEQ ID NO. 95, and SEQ ID NO. 114;

and a DVD light chain amino acid sequence selected from the group consisting of SEQ ID NO. 86, SEQ ID NO. 91, SEQ ID NO. 94, SEQ ID NO. 46, SEQ ID NO. 96, and SEQ ID NO. 116.

35. The binding protein according to claim 28 wherein the chemokine is selected from the group consisting of CCR2, CCR5 and CXCL-13.

36. The binding protein according to claim 28 wherein the cell surface protein is an integrin.

37. The binding protein according to claim 28 wherein the cell surface proteins are CD-20 and CD3.

38. The binding protein according to claim 37, wherein the binding protein comprises a DVD heavy chain amino acid sequence is SEQ ID NO. 97, and a DVD light chain SEQ ID NO. 101.

39. The binding protein according to claim 28 wherein the enzyme is selected from the group consisting of kinases and proteases.

40. The binding protein according to claim 28 wherein the receptor is selected from the group consisting of lymphokine receptor, monokine receptor, and polypeptide hormone receptor.

41. The binding protein according to claim 28, wherein said binding protein has an on rate constant (Kon) to said one or more targets selected from the group consisting of:

at least about 10²M<sup>−

1</sup>s<sup>−

1</sup>;

at least about 10³M<sup>−

1</sup>s<sup>−

1</sup>;

at least about 10⁴M<sup>−

1</sup>s<sup>−

1</sup>;

at least about 10⁵M<sup>−

1</sup>s<sup>−

1</sup>; and

at least about 10⁶M<sup>−

1</sup>s<sup>−

1</sup>, as measured by surface plasmon resonance.

42. The binding protein according to claim 28, wherein said binding protein has an off rate constant (Koff) to said one or more targets selected from the group consisting of:

at most about 10<sup>−

3</sup>s<sup>−

1</sup>;

at most about 10<sup>−

4</sup>s<sup>−

1</sup>;

at most about 10<sup>−

5</sup>s<sup>−

1</sup>; and

at most about 10<sup>−

6</sup>s<sup>−

1</sup>, as measured by surface plasmon resonance.

43. The binding protein according to claim 28, wherein said binding protein has a dissociation constant (K_D) to said one or more targets selected from the group consisting of:

at most about 10<sup>−

7</sup> M;

at most about 10<sup>−

8</sup> M;

at most about 10<sup>−

9</sup> M;

at most about 10<sup>−

10</sup> M;

at most about 10<sup>−

11</sup> M;

at most about 10<sup>−

12</sup> M; and

at most 10<sup>−

13</sup> M.

44. A binding protein conjugate comprising a binding protein described in any one of claims 1-43, said binding protein conjugate further comprising an agent selected from the group consisting of;

an immunoadhension molecule, an imaging agent, a therapeutic agent, and a cytotoxic agent.

45. The binding protein conjugate according to claim 44, wherein said agent is an imaging agent selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.

46. The binding protein conjugate according to claim 45, wherein said imaging agent is a radiolabel selected from the group consisting of:

³H, ¹⁴C, ³⁵S, ⁹⁰Y, ⁹⁹Tc, ¹¹¹In, ¹²⁵I, ¹¹³I, ¹⁷⁷Lu, ¹⁶⁶Ho, and ¹⁵³Sm.

47. The binding protein conjugate according to claim 45, wherein said agent is a therapeutic or cytotoxic agent selected from the group consisting of;

an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, toxin, and an apoptotic agent.

48. The binding protein according to claim 1, wherein said binding protein is a crystallized binding protein.

49. The crystallized binding protein according to claim 48, wherein said crystal is a carrier-free pharmaceutical controlled release crystal.

50. The crystallized binding protein according to claim 48, wherein said binding protein has a greater half life in vivo than the soluble counterpart of said binding protein.

51. The crystallized binding protein according to claim 48, wherein said binding protein retains biological activity.

52. An isolated nucleic acid encoding a binding protein amino acid sequence of any one of claims 1-22.

53. A vector comprising an isolated nucleic acid according to claim 52.

54. The vector of claim 53 wherein said vector is selected from the group consisting of pcDNA, pTT, pTT3, pEFBOS, pBV, pJV, pcDNA3.1 TOPO, pEF6 TOPO, and pBJ.

55. A host cell comprising a vector according to claim 53.

56. The host cell according to claim 55, wherein said host cell is a prokaryotic cell.

57. The host cell according to claim 56, wherein said host cell is E. Coli.

58. The host cell according to claim 55, wherein said host cell is a eukaryotic cell.

59. The host cell according to claim 58, wherein said eukaryotic cell is selected from the group consisting of protist cell, animal cell, plant cell and fungal cell.

60. The host cell according to claim 58, wherein said eukaryotic cell is an animal cell selected from the group consisting of;

a mammalian cell, an avian cell, and an insect cell.

61. The host cell according to claim 58, wherein said host cell is a CHO cell.

62. The host cell according to claim 58, wherein said host cell is COS.

63. The host cell according to claim 58, wherein said host cell is a yeast cell.

64. The host cell according to claim 63, wherein said yeast cell is Saccharomyces cerevisiae.

65. The host cell according to claim 58, wherein said host cell is an insect Sf9 cell.

66. A method of producing a binding protein, comprising culturing a host cell described in any one of claims 55-65 in culture medium under conditions sufficient to produce the binding protein.

67. The method of claim 66, wherein 50%-75% of the binding protein produced is a dual specific tetravalent binding protein.

68. The method of claim 66, wherein 75%-90% of the binding protein produced is a dual specific tetravalent binding protein.

69. The method of claim 66, wherein 90%-95% of the binding protein produced is a dual specific tetravalent binding protein.

70. A protein produced according to the method of claim 66.

71. A pharmaceutical composition comprising the binding protein of any one of claims 1-51 and 70, and a pharmaceutically acceptable carrier.

72. The pharmaceutical composition of claim 71 further comprising at least one additional therapeutic agent.

73. The pharmaceutical composition of claim 72, wherein said additional agent is selected from the group consisting of:

Therapeutic agent, imaging agent, cytotoxic agent, angiogenesis inhibitors;

kinase inhibitors;

co-stimulation molecule blockers;

adhesion molecule blockers;

anti-cytokine antibody or functional fragment thereof;

methotrexate;

cyclosporin;

rapamycin;

FK506;

detectable label or reporter;

a TNF antagonist;

an antirheumatic;

a muscle relaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteriod, an anabolic steroid, an erythropoietin, an immunization, an immunoglobulin, an immunosuppressive, a growth hormone, a hormone replacement drug, a radiopharmaceutical, an antidepressant, an antipsychotic, a stimulant, an asthma medication, a beta agonist, an inhaled steroid, an epinephrine or analog, a cytokine, and a cytokine antagonist.

74. A method for treating a subject for a disease or a disorder by administering to the subject the binding protein of any one of claims 1-51 and 70 such that treatment is achieved.

75. The method of claim 74, wherein said disorder is selected from the group comprising rheumatoid arthritis, osteoarthritis, juvenile chronic arthritis, septic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, spondyloarthropathy, systemic lupus erythematosus, Crohn'"'"'s disease, ulcerative colitis, inflammatory bowel disease, insulin dependent diabetes mellitus, thyroiditis, asthma, allergic diseases, psoriasis, dermatitis scleroderma, graft versus host disease, organ transplant rejection, acute or chronic immune disease associated with organ transplantation, sarcoidosis, atherosclerosis, disseminated intravascular coagulation, Kawasaki'"'"'s disease, Grave'"'"'s disease, nephrotic syndrome, chronic fatigue syndrome, Wegener'"'"'s granulomatosis, Henoch-Schoenlein purpurea, microscopic vasculitis of the kidneys, chronic active hepatitis, uveitis, septic shock, toxic shock syndrome, sepsis syndrome, cachexia, infectious diseases, parasitic diseases, acquired immunodeficiency syndrome, acute transverse myelitis, Huntington'"'"'s chorea, Parkinson'"'"'s disease, Alzheimer'"'"'s disease, stroke, primary biliary cirrhosis, hemolytic anemia, malignancies, heart failure, myocardial infarction, Addison'"'"'s disease, sporadic, polyglandular deficiency type I and polyglandular deficiency type II, Schmidt'"'"'s syndrome, adult (acute) respiratory distress syndrome, alopecia, alopecia areata, seronegative arthopathy, arthropathy, Reiter'"'"'s disease, psoriatic arthropathy, ulcerative colitic arthropathy, enteropathic synovitis, chlamydia, yersinia and salmonella associated arthropathy, spondyloarthopathy, atheromatous disease/arteriosclerosis, atopic allergy, autoimmune bullous disease, pemphigus vulgaris, pemphigus foliaceus, pemphigoid, linear IgA disease, autoimmune haemolytic anaemia, Coombs positive haemolytic anaemia, acquired pernicious anaemia, juvenile pernicious anaemia, myalgic encephalitis/Royal Free Disease, chronic mucocutaneous candidiasis, giant cell arteritis, primary sć

lerosing hepatitis, cryptogenic autoimmune hepatitis, Acquired Immunodeficiency Disease Syndrome, Acquired Immunodeficiency Related Diseases, Hepatitis B, Hepatitis C, common varied immunodeficiency (common variable hypogammaglobulinaemia), dilated cardiomyopathy, female infertility, ovarian failure, premature ovarian failure, fibrotic lung disease, cryptogenic fibrosing alveolitis, post-inflammatory interstitial lung disease, interstitial pneumonitis, connective tissue disease associated interstitial lung disease, mixed connective tissue disease associated lung disease, systemic sclerosis associated interstitial lung disease, rheumatoid arthritis associated interstitial lung disease, systemic lupus erythematosus associated lung disease, dermatomyositis/polymyositis associated lung disease, Sjö

gren'"'"'s disease associated lung disease, ankylosing spondylitis associated lung disease, vasculitic diffuse lung disease, haemosiderosis associated lung disease, drug-induced interstitial lung disease, fibrosis, radiation fibrosis, bronchiolitis obliterans, chronic eosinophilic pneumonia, lymphocytic infiltrative lung disease, postinfectious interstitial lung disease, gouty arthritis, autoimmune hepatitis, type-1 autoimmune hepatitis (classical autoimmune or lupoid hepatitis), type-2 autoimmune hepatitis (anti-LKM antibody hepatitis), autoimmune mediated hypoglycaemia, type B insulin resistance with acanthosis nigricans, hypoparathyroidism, acute immune disease associated with organ transplantation, chronic immune disease associated with organ transplantation, osteoarthrosis, primary sclerosing cholangitis, psoriasis type 1, psoriasis type 2, idiopathic leucopaenia, autoimmune neutropaenia, renal disease NOS, glomerulonephritides, microscopic vasulitis of the kidneys, lyme disease, discoid lupus erythematosus, male infertility idiopathic or NOS, sperm autoimmunity, multiple sclerosis (all subtypes), sympathetic ophthalmia, pulmonary hypertension secondary to connective tissue disease, Goodpasture'"'"'s syndrome, pulmonary manifestation of polyarteritis nodosa, acute rheumatic fever, rheumatoid spondylitis, Still'"'"'s disease, systemic sclerosis, Sjö

rgren'"'"'s syndrome, Takayasu'"'"'s disease/arteritis, autoimmune thrombocytopaenia, idiopathic thrombocytopaenia, autoimmune thyroid disease, hyperthyroidism, goitrous autoimmune hypothyroidism (Hashimoto'"'"'s disease), atrophic autoimmune hypothyroidism, primary myxoedema, phacogenic uveitis, primary vasculitis, vitiligo acute liver disease, chronic liver diseases, alcoholic cirrhosis, alcohol-induced liver injury, choleosatatis, idiosyncratic liver disease, Drug-Induced hepatitis, Non-alcoholic Steatohepatitis, allergy and asthma, group B streptococci (GBS) infection, mental disorders (e.g., depression and schizophrenia), Th2 Type and Th1 Type mediated diseases, acute and chronic pain (different forms of pain), and cancers such as lung, breast, stomach, bladder, colon, pancreas, ovarian, prostate and rectal cancer and hematopoietic malignancies (leukemia and lymphoma) Abetalipoprotemia, Acrocyanosis, acute and chronic parasitic or infectious processes, acute leukemia, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute or chronic bacterial infection, acute pancreatitis, acute renal failure, adenocarcinomas, aerial ectopic beats, AIDS dementia complex, alcohol-induced hepatitis, allergic conjunctivitis, allergic contact dermatitis, allergic rhinitis, allograft rejection, alpha-1-antitrypsin deficiency, amyotrophic lateral sclerosis, anemia, angina pectoris, anterior hom cell degeneration, anti cd3 therapy, antiphospholipid syndrome, anti-receptor hypersensitivity reactions, aordic and peripheral aneuryisms, aortic dissection, arterial hypertension, arteriosclerosis, arteriovenous fistula, ataxia, atrial fibrillation (sustained or paroxysmal), atrial flutter, atrioventricular block, B cell lymphoma, bone graft rejection, bone marrow transplant (BMT) rejection, bundle branch block, Burkitt'"'"'s lymphoma, Burns, cardiac arrhythmias, cardiac stun syndrome, cardiac tumors, cardiomyopathy, cardiopulmonary bypass inflammation response, cartilage transplant rejection, cerebellar cortical degenerations, cerebellar disorders, chaotic or multifocal atrial tachycardia, chemotherapy associated disorders, chromic myelocytic leukemia (CML), chronic alcoholism, chronic inflammatory pathologies, chronic lymphocytic leukemia (CLL), chronic obstructive pulmonary disease (COPD), chronic salicylate intoxication, colorectal carcinoma, congestive heart failure, conjunctivitis, contact dermatitis, cor pulmonale, coronary artery disease, Creutzfeldt-Jakob disease, culture negative sepsis, cystic fibrosis, cytokine therapy associated disorders, Dementia pugilistica, demyelinating diseases, dengue hemorrhagic fever, dermatitis, dermatologic conditions, diabetes, diabetes mellitus, diabetic ateriosclerotic disease, Diffuse Lewy body disease, dilated congestive cardiomyopathy, disorders of the basal ganglia, Down'"'"'s Syndrome in middle age, drug-induced movement disorders induced by drugs which block CNS dopamine receptors, drug sensitivity, eczema, encephalomyelitis, endocarditis, endocrinopathy, epiglottitis, epstein-barr virus infection, erythromelalgia, extrapyramidal and cerebellar disorders, familial hematophagocytic lymphohistiocytosis, fetal thymus implant rejection, Friedreich'"'"'s ataxia, functional peripheral arterial disorders, fungal sepsis, gas gangrene, gastric ulcer, glomerular nephritis, graft rejection of any organ or tissue, gram negative sepsis, gram positive sepsis, granulomas due to intracellular organisms, hairy cell leukemia, Hallerrorden-Spatz disease, hashimoto'"'"'s thyroiditis, hay fever, heart transplant rejection, hemachromatosis, hemodialysis, hemolytic uremic syndrome/thrombolytic thrombocytopenic purpura, hemorrhage, hepatitis (A), His bundle arrythmias, HIV infection/HIV neuropathy, Hodgkin'"'"'s disease, hyperkinetic movement disorders, hypersensitity reactions, hypersensitivity pneumonitis, hypertension, hypokinetic movement disorders, hypothalamic-pituitary-adrenal axis evaluation, idiopathic Addison'"'"'s disease, idiopathic pulmonary fibrosis, antibody mediated cytotoxicity, Asthenia, infantile spinal muscular atrophy, inflammation of the aorta, influenza a, ionizing radiation exposure, iridocyclitis/uveitis/optic neuritis, ischemia-reperfusion injury, ischemic stroke, juvenile rheumatoid arthritis, juvenile spinal muscular atrophy, Kaposi'"'"'s sarcoma, kidney transplant rejection, legionella, leishmaniasis, leprosy, lesions of the corticospinal system, lipedema, liver transplant rejection, lymphederma, malaria, malignant Lymphoma, malignant histiocytosis, malignant melanoma, meningitis, meningococcemia, metabolic/idiopathic, migraine headache, mitochondrial multi.system disorder, mixed connective tissue disease, monoclonal gammopathy, multiple myeloma, multiple systems degenerations (Mencel Dejerine-Thomas Shi-Drager and Machado-Joseph), myasthenia gravis, mycobacterium avium intracellulare, mycobacterium tuberculosis, myelodyplastic syndrome, myocardial infarction, myocardial ischemic disorders, nasopharyngeal carcinoma, neonatal chronic lung disease, nephritis, nephrosis, neurodegenerative diseases, neurogenic I muscular atrophies, neutropenic fever, non-hodgkins lymphoma, occlusion of the abdominal aorta and its branches, occulsive arterial disorders, okt3 therapy, orchitis/epidydimitis, orchitis/vasectomy reversal procedures, organomegaly, osteoporosis, pancreas transplant rejection, pancreatic carcinoma, paraneoplastic syndrome/hypercalcemia of malignancy, parathyroid transplant rejection, pelvic inflammatory disease, perennial rhinitis, pericardial disease, peripheral atherlosclerotic disease, peripheral vascular disorders, peritonitis, pernicious anemia, pneumocystis carinii pneumonia, pneumonia, POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes syndrome), post perfusion syndrome, post pump syndrome, post-MI cardiotomy syndrome, preeclampsia, Progressive supranucleo Palsy, primary pulmonary hypertension, radiation therapy, Raynaud'"'"'s phenomenon and disease, Raynoud'"'"'s disease, Refsum'"'"'s disease, regular narrow QRS tachycardia, renovascular hypertension, reperfusion injury, restrictive cardiomyopathy, sarcomas, scleroderma, senile chorea, Senile Dementia of Lewy body type, seronegative arthropathies, shock, sickle cell anemia, skin allograft rejection, skin changes syndrome, small bowel transplant rejection, solid tumors, specific arrythmias, spinal ataxia, spinocerebellar degenerations, streptococcal myositis, structural lesions of the cerebellum, Subacute sclerosing panencephalitis, Syncope, syphilis of the cardiovascular system, systemic anaphalaxis, systemic inflammatory response syndrome, systemic onset juvenile rheumatoid arthritis, T-cell or FAB ALL, Telangiectasia, thromboangitis obliterans, thrombocytopenia, toxicity, transplants, trauma/hemorrhage, type III hypersensitivity reactions, type IV hypersensitivity, unstable angina, uremia, urosepsis, urticaria, valvular heart diseases, varicose veins, vasculitis, venous diseases, venous thrombosis, ventricular fibrillation, viral and fungal infections, vital encephalitis/aseptic meningitis, vital-associated hemaphagocytic syndrome, Wernicke-Korsakoff syndrome, Wilson'"'"'s disease, xenograft rejection of any organ or tissue.

76. The method according to claim 74, wherein said administering to the subject is by at least one mode selected from parenteral, subcutaneous, intramuscular, intravenous, intrarticular, intrabronchial, intraabdominal, intracapsular, intracartilaginous, intracavitary, intracelial, intracerebellar, intracerebroventricular, intracolic, intracervical, intragastric, intrahepatic, intramyocardial, intraosteal, intrapelvic, intrapericardiac, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal, intrasynovial, intrathoracic, intrauterine, intravesical, bolus, vaginal, rectal, buccal, sublingual, intranasal, and transdermal.

11. A binding protein comprising a polypeptide chain, wherein said polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable domain, VD2 is a second heavy chain variable domain, C is a heavy chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 is an Fc region.

20. A binding protein comprising a polypeptide chain, wherein said polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain variable domain, VD2 is a second light chain variable domain, C is a light chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 does not comprise an Fc region.

21. A binding protein comprising first and second polypeptide chains, wherein said first polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable domain, VD2 is a second heavy chain variable domain, C is a heavy chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 is an Fc region;

and said second polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain variable domain, VD2 is a second light chain variable domain, C is a light chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 does not comprise an Fc region.

22. A binding protein comprising four polypeptide chains, wherein two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable domain, VD2 is a second heavy chain variable domain, C is a heavy chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 is an Fc region;

and two polypeptide chains comprises VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain variable domain, VD2 is a second light chain variable domain, C is a light chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 does not comprise an Fc region.