CN117917436A

CN117917436A - Antibody capable of specifically recognizing TRAIL and application thereof

Info

Publication number: CN117917436A
Application number: CN202311317909.5A
Authority: CN
Inventors: 王广菲; 任晓叶; 张思
Original assignee: Beijing Solobio Genetechnology Co Ltd
Current assignee: Beijing Solobio Genetechnology Co Ltd
Priority date: 2022-10-20
Filing date: 2023-10-12
Publication date: 2024-04-23

Abstract

The application relates to an antibody or antigen binding fragment specifically recognizing TRAIL, a preparation method and application thereof.

Description

Antibody capable of specifically recognizing TRAIL and application thereof

Cross Reference to Related Applications

The present application claims priority to PCT application No. PCT/CN2022/126334, filing date 2022.10.20, entitled "antibodies specifically recognizing TRAIL and uses thereof", and the entire contents of which are incorporated herein by reference.

Reference to an electronic sequence Listing

The contents of the electronic sequence Listing (text name: CN_202207067466_SEQLIST. Xml, recording date: 2023.09.14, size: 82 KB) are incorporated herein by reference in their entirety.

Technical Field

Background

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a type II transmembrane protein of the TNF superfamily that can be cleaved from the cell surface by cysteine proteases and matrix metalloproteinase 2 (MMP-2), yielding a soluble form (Wiley,S R et al.Immunity vol.3,6(1995):673-82.;Mariani,S M,and P H Krammer.European journal of immunology vol.28,3(1998):973-82.).TRAIL that remains pro-apoptotic, which is constitutively present in a variety of tissues but is expressed primarily in Natural Killer (NK) cells and macrophages (Wang, s.current MEDICINAL CHEMISTRY vol.17,29 (2010): 3309-17.).

TRAIL has 5 different receptors: TRAIL-R1, TRAIL-R2, dcR1, dcR2 and osteoprotegerin. Among them, TRAIL-R1 (also called DR 4) and TRAIL-R2 (also called DR 5) contain cytoplasmic regions called death domains, which allow the receptor to bind to cognate ligands and trigger cytotoxic signals. DcR1 does not contain a death domain, dcR2 contains an incomplete death domain, and osteoprotegerin is a soluble TRAIL receptor. These three receptors can act as decoy receptors, sequester TRAIL binding to its pro-apoptotic receptor or interfere with death receptor-mediated death signaling, thereby avoiding the production of apoptosis (Walczak, H, and P H kramer. Experimental CELL RESEARCH vol.256,1 (2000): 58-66.). DR4 is typically expressed in most human tissues, including spleen, thymus, liver, peripheral blood leukocytes, activated T cells, and small intestine. Expression of DR5 is ubiquitous in normal and tumor tissues, but is particularly abundant in spleen, peripheral blood leukocytes and activated lymphocytes (Wang, s.current MEDICINAL CHEMISTRY vol.17,29 (2010): 3309-17.).

TRAIL, upon binding to death receptor DR4 or DR5, induces receptor trimerization and recruits Fas-associated protein (FADD) and membrane proximal caspases (caspases 8 or 10) to form a receptor complex called death-inducing signaling complex (DISC). Caspases are synthesized in inactive zymogen forms that induce their own activation upon recruitment to form the DISC. The subsequently activated caspases cut directly downstream effector caspases (caspases 3,6 and 7), resulting in their activation. Activated caspase 3 cleaves a variety of cellular proteins leading to apoptosis (Ashkenazi, A, and V M Dixit. Science (New York, N.Y.) vol.281,5381 (1998): 1305-8.). In type I cells, a relatively large number of DISC assemble and internalize rapidly, producing enough caspase 8 (or 10) and signaling to promote cell death. In contrast, DISC formation is delayed and less efficient in type II cells. Thus, activation of caspase 8 in type II cells is insufficient to activate apoptosis. The execution of apoptosis requires that mitochondria mediate cleavage of BH3 interaction domain death agonists (Bid) by caspase 8 to produce truncated forms of Bid (tBid), which further induce activation of Bax and Bak in mitochondria. Activated Bax and Bak promote loss of mitochondrial membrane potential and release of cytochrome c, which in turn leads to apoptosis (Barnhart,Bryan C et al.Seminars in immunology vol.15,3(2003):185-93.;Kohlhaas,Susan L et al.The Journal of biological chemistry vol.282,17(2007):12831-41.).

In addition to being able to induce cell death, TRAIL signaling may also promote cell development, survival and proliferation (Lin,Y et al.Molecular and cellular biology vol.20,18(2000):6638-45.;Lee,Tae-Jin et al.Biochemical and biophysical research communications vol.351,4(2006):1024-30.). by activating nuclear factors- κb (NF-kB), MAP kinase (MAPK) and Akt, e.g., TRAIL may trigger the formation of cytoplasmic complexes comprising FADD, TNF receptor-related factor 2 (TRAF 2) and NF- κb essential modifier (NEMO), which may lead to activation of other signaling pathways, including activation of NF- κb and MAPK, and production of pro-inflammatory cytokines and chemokines.

Studies have shown that TRAIL plays a role not only in tumors but also in the development and progression of various diseases such as autoimmune diseases, liver, kidney, graft rejection, spinal cord injury, alzheimer's disease, etc. (Yin,Xiao-Ming,and Wen-Xing Ding.Current molecular medicine vol.3,6(2003):491-508.;Zhang,Yun et al.Immunology letters vol.152,1(2013):1-7.;Burgaletto,Chiara et al.Journal of neuroinflammation vol.17,1 298.13Oct.2020).Hameed et al have shown that in toxin-induced PAH rat models, TRAIL expression in pulmonary endothelial and epithelial cells is enhanced at peak vascular remodeling and elevated right ventricular blood pressure, and administration of TRAIL-antagonistic antibodies can significantly reduce pulmonary arterial hypertension (Hameed, abdul G et al, the Journal of experimental medicine vol.209,11 (2012): 1919-35.). Studies by Ochi et al have shown that liver NK cells expressing TRAIL are toxic to mice own hepatocytes, and that anti-TRAIL antibodies partially inhibit their hepatotoxicity (Ochi, makoto et al. Hepatology (Baltimore, md.) vol.39,5 (2004): 1321-31.). Therefore, the development of inhibitors against TRAIL targets is of great importance.

Currently, fusion proteins sDR5-Fc with the function of blocking TRAIL-DR5 signaling pathway have been reported for TRAIL inhibitors, for example, as disclosed in China patent CN 108997503B. However, no monoclonal antibody drug targeting TRAIL has been marketed, and therefore development of a monoclonal antibody drug targeting TRAIL is needed.

The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are incorporated by reference in their entirety.

Summary of the application

In some embodiments, an isolated anti-TRAIL antibody is provided comprising: (i) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 50V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 70; (ii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 51V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 71; (iii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 52V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 72; (iv) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 53V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 73; (v) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 54V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 74; (vi) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 55V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 75; (vii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 56V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 76; (viii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 57V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 74; (ix) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 58V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 77; (x) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID NO 59V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 78; (xi) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 60V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 79; (xii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 61V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 80; (xiii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 62V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 81; (xiv) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 63V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 82; (xv) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 64V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 of V _L as shown in amino acid sequence SEQ ID NO 83; (xvi) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 65V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 84; (xvii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 66, which V _H comprises; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 85; (xviii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 67V _H; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 86; (xix) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by V _H as shown in amino acid sequence SEQ ID NO 68; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 87; or (xx) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 69; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 88.

In some embodiments, an isolated anti-TRAIL antibody is provided comprising: (i) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 29, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (ii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 30, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (iii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 31, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (iv) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (v) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (vi) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:20, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (vii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 21, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (viii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 22, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising an amino acid sequence

32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (ix) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 33, LC-CDR2 comprising the amino acid sequence SEQ ID No. 39, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 43, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (x) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 7, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 34, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 44, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xi) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 44, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 44, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xiii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and

V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 45, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xiv) V _H, the V _H comprising: HC-CDR1 comprising an amino acid sequence

8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 35, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 45, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xv) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 45, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xvi) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 46, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xvii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 26, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 36, LC-CDR2 comprising the amino acid sequence SEQ ID No. 40, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 47, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xviii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 48, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; (xix) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 27, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 44, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; or (xx) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:49, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, any of the isolated anti-TRAIL antibodies described above, comprising: (i) V _H comprising the amino acid sequence set forth in SEQ ID No. 50 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 50; and V _L comprising the amino acid sequence set forth in SEQ ID No. 70 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 70; (ii) V _H comprising the amino acid sequence set forth in SEQ ID No. 51 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 51; and V _L comprising the amino acid sequence set forth in SEQ ID No. 71 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 71; (iii) V _H comprising the amino acid sequence set forth in SEQ ID No. 52 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 52; and V _L comprising the amino acid sequence set forth in SEQ ID No. 72 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 72; (iv) V _H comprising the amino acid sequence set forth in SEQ ID No. 53 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 53; and V _L comprising the amino acid sequence set forth in SEQ ID No. 73 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 73; (v) V _H comprising the amino acid sequence set forth in SEQ ID No. 54 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 54; and V _L comprising the amino acid sequence set forth in SEQ ID No. 74 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 74; (vi) V _H comprising the amino acid sequence set forth in SEQ ID No. 55 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 55; and V _L comprising the amino acid sequence set forth in SEQ ID No. 75 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 75; (vii) V _H comprising the amino acid sequence set forth in SEQ ID No. 56 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 56; and V _L comprising the amino acid sequence set forth in SEQ ID No. 76 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 76; (viii) V _H comprising the amino acid sequence set forth in SEQ ID No. 57 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 57; and V _L which comprises

An amino acid sequence set forth in SEQ ID NO. 74 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 74; (ix) V _H comprising the amino acid sequence shown in SEQ ID NO. 58 or a variant thereof, said variant being identical to

The amino acid sequence shown in SEQ ID NO. 58 has at least about 80% sequence identity; and V _L comprising the amino acid sequence set forth in SEQ ID No. 77 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 77; (x) V _H comprising the amino acid sequence set forth in SEQ ID No. 59 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 59; and V _L comprising the amino acid sequence set forth in SEQ ID No. 78 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 78; (xi) V _H comprising the amino acid sequence set forth in SEQ ID No. 60 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 60; and V _L comprising the amino acid sequence set forth in SEQ ID No. 79 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 79; (xii) V _H comprising the amino acid sequence set forth in SEQ ID No. 61 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 61; and V _L comprising the amino acid sequence set forth in SEQ ID No. 80 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 80; (xiii) V _H comprising the amino acid sequence set forth in SEQ ID No. 62 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 62; and V _L comprising the amino acid sequence set forth in SEQ ID No. 81 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 81; (xiv) V _H comprising the amino acid sequence set forth in SEQ ID No. 63 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 63; and V _L comprising the amino acid sequence set forth in SEQ ID No. 82 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 82; (xv) V _H comprising the amino acid sequence set forth in SEQ ID No. 64 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 64; and V _L comprising the amino acid sequence shown as SEQ ID NO. 83 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown as SEQ ID NO. 83;

(xvi) V _H comprising the amino acid sequence set forth in SEQ ID No. 65 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 65; and V _L comprising the amino acid sequence set forth in SEQ ID No. 84 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 84; (xvii) V _H comprising the amino acid sequence set forth in SEQ ID No. 66 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 66; and V _L comprising the amino acid sequence set forth in SEQ ID No. 85 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 85; (xviii) V _H comprising the amino acid sequence set forth in SEQ ID No. 67 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 67; and V _L comprising the amino acid sequence set forth in SEQ ID No. 86 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 86; (xix) V _H comprising the amino acid sequence set forth in SEQ ID No. 68 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 68; and

V _L comprising the amino acid sequence set forth in SEQ ID No. 87 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 87; or (xx) V _H comprising the amino acid sequence set forth in SEQ ID NO. 69, or a variant thereof, which has at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 69; and V _L comprising the amino acid sequence shown as SEQ ID NO. 88 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown as SEQ ID NO. 88.

In some embodiments, an isolated anti-TRAIL antibody is provided that competes with any of the isolated anti-TRAIL antibodies described above for specific binding to TRAIL. In some embodiments, an isolated anti-TRAIL antibody is provided that specifically binds to the same epitope as any of the isolated anti-TRAIL antibodies described above.

In some embodiments, any of the isolated anti-TRAIL antibodies described above, comprising an Fc fragment. In some embodiments, the isolated anti-TRAIL antibody is a full-length IgG antibody. In some embodiments, the isolated anti-TRAIL antibody is a full length IgG1, igG2, igG3, or IgG4 antibody. In some embodiments, the isolated anti-TRAIL antibody is a chimeric, fully human or humanized antibody. In some embodiments, the isolated anti-TRAIL antibody is an antigen-binding fragment selected from the group consisting of Fab, fab ', F (ab) ' ₂, fab ' -SH, single chain Fv (scFv), fv fragment, dAb, fd, nanobody (nanobody), diabody (diabody), and linear antibody.

In some embodiments, an isolated nucleic acid molecule encoding any one of the anti-TRAIL antibodies described above is provided. In some embodiments, a vector is provided, the vector comprising any one of the nucleic acid molecules described above. In some embodiments, a host cell is provided, the host cell comprising any one of the anti-TRAIL antibodies described above, any one of the nucleic acid molecules described above, or any one of the vectors described above. In some embodiments, a method of making an anti-TRAIL antibody is provided, comprising: a) Culturing any of the above host cells under conditions effective to express an anti-TRAIL antibody; and b) obtaining the expressed anti-TRAIL antibody from the host cell.

In some embodiments, there is provided a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of any one of the anti-TRAIL antibodies described above. In some embodiments, there is provided the use of any one of the anti-TRAIL antibodies described above in the manufacture of a pharmaceutical composition for treating a disease or condition in a subject in need thereof. In some embodiments, there is provided the use of any one of the anti-TRAIL antibodies or a pharmaceutical composition comprising an anti-TRAIL antibody as described above in the manufacture of a medicament for the treatment of a disease or disorder. In some embodiments, the disease or disorder is associated with TRAIL signaling, including inflammatory diseases, autoimmune diseases, transplantation-related diseases, liver diseases, neurodegenerative diseases, or cancers or disorders. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer.

Also provided are pharmaceutical compositions, kits, and articles of manufacture comprising any of the anti-TRAIL antibodies described above.

Detailed description of the application

In one aspect, the application provides anti-TRAIL antibody molecules. Through scFv phage library screening in combination with appropriately designed biochemical and biological experiments, highly potent antibody molecules have been identified that are capable of binding human TRAIL and inhibiting the interaction of human TRAIL with its receptor. The results presented herein demonstrate that the antibodies of the application have better biological activity than the commercial anti-TRAIL antibody MAB 375-500.

Anti-TRAIL antibodies provided herein include, for example, full length anti-TRAIL antibodies, anti-TRAIL single chain antibodies (scFvs), anti-TRAIL Fc fusion proteins, multi-specific (e.g., bispecific) anti-TRAIL antibodies, anti-TRAIL immunoconjugates, and the like.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 29, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 30, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 31, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:20, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 21, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 22, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 33, LC-CDR2 comprising the amino acid sequence SEQ ID No. 39, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 43, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 7, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO 34, LC-CDR2 comprising the amino acid sequence SEQ ID NO 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:35, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 46, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, and

HC-CDR3 comprising the amino acid sequence SEQ ID NO. 26, or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:36, LC-CDR2 comprising the amino acid sequence SEQ ID NO:40, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:47, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 48, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 27, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In another aspect, the application provides an anti-TRAIL antibody comprising: v _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, and

A HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28, or a variant of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:49, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

Also provided are nucleic acids encoding anti-TRAIL antibodies, compositions comprising anti-TRAIL antibodies, and methods of making and using anti-TRAIL antibodies.

Definition of the definition

As used herein, a "treatment" or "treatment" is a method of achieving a beneficial or desired result, including clinical results. In view of the objects of the present application, such beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviating one or more symptoms caused by the disease, alleviating the extent of the disease, stabilizing the disease (e.g., preventing or delaying the progression of the disease), preventing or delaying the spread of the disease, preventing or delaying the recurrence of the disease, delaying or slowing the progression of the disease, ameliorating the disease state, alleviating the disease (partially or wholly), reducing the dosage of one or more other drugs required to treat the disease, delaying the progression of the disease, improving or enhancing the quality of life, increasing weight, and/or prolonging survival. Also, "treatment" includes reduction of disease pathology results. The methods of the present application contemplate any one or more aspects of these treatments.

The term "antibody" includes full length antibodies and antigen binding fragments thereof. Full length antibodies include two heavy chains and two light chains. The variable regions of the light and heavy chains are responsible for antigen binding. The variable region in both chains typically comprises 3 hypervariable loops, known as Complementarity Determining Regions (CDRs) (light chain (LC) CDRs comprise LC-CDR1, LC-CDR2 and LC-CDR3, and Heavy Chain (HC) CDRs comprise HC-CDR1, HC-CDR2 and HC-CDR 3). CDR boundaries of antibodies or antigen binding fragments disclosed herein may be defined or identified by Kabat, chothia or Al-Lazikani conventions (Al-Lazikani 1997;Chothia 1985;Chothia 1987;Chothia1989;Kabat 1987;Kabat 1991). The 3 CDR regions of the heavy or light chain are inserted between flanking segments called Framework Regions (FRs) which are more conserved than the CDR regions and form a scaffold supporting the hypervariable loops. The constant regions of the heavy and light chains are not involved in antigen binding, but exhibit multiple effector functions. Antibodies are classified based on the amino acid sequence of their heavy chain constant regions. The five main classes or isotypes of antibodies are IgA, igD, igE, igG and IgM, which are characterized by having alpha, delta, epsilon, gamma, and mu heavy chains, respectively. Several major antibody classes are classified into subclasses, such as IgG1 (gamma 1 heavy chain), igG2 (gamma 2 heavy chain), igG3 (gamma 3 heavy chain), igG4 (gamma 4 heavy chain), igA1 (alpha 1 heavy chain), or IgA2 (alpha 2 heavy chain).

As used herein, the term "antigen-binding fragment" includes antibody fragments, e.g., diabodies (diabodies), fab ', F (ab ') ₂, fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv) ₂, bispecific dsFv (dsFv-dsFv '), disulfide-stabilized diabodies (ds diabodies), single chain Fv (scFv), scFv dimers (diabodies), multispecific antibodies consisting of antibody fragments comprising one or more CDRs, single domain antibodies, nanobodies (nanobodies), domain antibodies, bivalent domain antibodies, or any other antibody fragment capable of binding an antigen but not comprising an intact antibody structure. The antigen binding fragment is capable of binding the same antigen as the parent antibody or parent antibody fragment (e.g., parent scFv). Antigen binding fragments also include fusion proteins comprising the above antibody fragments. In some embodiments, an antigen binding fragment may include one or more CDRs from a particular human antibody grafted to a framework region from one or more different human antibodies.

As used herein, the term "epitope" refers to a specific group of atoms or amino acids on an antigen to which an antibody or antibody portion binds. If two antibodies or antibody portions exhibit competitive binding to an antigen, they may bind to the same epitope on the antigen.

As used herein, a first antibody "competes" with a second antibody for binding to a TRAIL target when the first antibody inhibits the second antibody from binding to the TRAIL target by at least 50% (e.g., at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) at an equimolar concentration, and vice versa. PCT publication WO 03/48731 describes a high throughput antibody "epitope categorization" method based on cross-competition.

As used herein, the term "specifically binds," "specifically recognizes," or "specific for," refers to a measurable and reproducible interaction, e.g., binding of an antibody to a target can determine the presence of the target in a heterogeneous population of molecules, including biomolecules. For example, an antibody being able to specifically recognize a target (which may be an epitope) means that the antibody binds to the target with a higher affinity, avidity, easier and/or longer lasting than other targets. In some embodiments, an antibody that specifically recognizes an antigen reacts with one or more antigenic determinants of the antigen with a binding affinity that is at least 10-fold greater than its binding affinity to other targets.

As used herein, an "isolated" anti-TRAIL antibody refers to an anti-TRAIL antibody that (1) is not associated with naturally occurring proteins, (2) does not contain other proteins of the same origin, (3) is expressed by cells of different species, or (4) is not found in nature.

As used herein, the term "isolated nucleic acid" refers to nucleic acids of genomic, cDNA, or synthetic origin, or a combination thereof. According to its origin, the term "isolated nucleic acid" means (1) not related to all or part of the polynucleotide found in nature in "isolated nucleic acid" (2) operably linked to a polynucleotide not linked thereto in nature, or (3) not present in nature as part of a longer sequence.

As used herein, the term "CDR" or "complementarity determining region" means a discontinuous antigen binding site found within the variable domains of heavy and light chain polypeptides. These particular regions have been described in literature Kabat et al.,J.Biol.Chem.252:6609-6616(1977);Kabat et al.,U.S.Dept.of Health and Human Services,"Sequences of proteins of immunological interest"(1991);Chothia et al.,J.Mol.Biol.196:901-917(1987);Al-Lazikani B.et al.,J.Mol.Biol.,273:927-948(1997);MacCallum et al.,J.Mol.Biol.262:732-745(1996);Abhinandan and Martin,Mol.Immunol.,45:3832-3839(2008);Lefranc M.P.et al.,Dev.Comp.Immunol.,27:55-77(2003); and honeygger and plackthun, j.mol.biol.,309:657-670 (2001), wherein these definitions include the coincidence or subset of amino acid residues when compared to each other. However, any definition of a CDR for an antibody or grafted antibody or variant thereof is intended to be included within the terms defined and used herein. The positions of the amino acid residues comprised by the CDRs defined by the various references cited above are listed in table 1 to illustrate the comparison. Algorithms and binding interfaces for CDR prediction are known in the art and include, for example, Abhinandan and Martin,Mol.Immunol.,45:3832-3839(2008);Ehrenmann F.et al.,Nucleic Acids Res.,38:D301-D307(2010); and Adolf-Bryfogle j.et al, nucleic Acids res, 43:d432-D438 (2015) are described. The content of the references cited in this paragraph is hereby incorporated by reference in its entirety for the purposes of the present application and possibly in one or more of the claims contained herein.

TABLE 1 CDR definition

	Kabat¹	Chothia²	MacCallum³	IMGT⁴	AHo⁵
						V_H CDR1	31-35	26-32	30-35	27-38	25-40
V_H CDR2	50-65	53-55	47-58	56-65	58-77
						V_H CDR3	95-102	96-101	93-101	105-117	109-137
V_L CDR1	24-34	26-32	30-36	27-38	25-40
						V_L CDR2	50-56	50-52	46-55	56-65	58-77
V_L CDR3	89-97	91-96	89-96	105-117	109-137

¹ Amino acid residue numbering refers to the nomenclature used in Kabat et al, supra

² Amino acid residue numbering refers to the nomenclature used in Chothia et al, supra

³ Amino acid residue numbering refers to the nomenclature method in MacCallum et al above

⁴ Amino acid residue numbering refers to the nomenclature method in LEFRANC ET al above

⁵ Amino acid residue numbering refers to the naming method in Honygger and Pluckthun, supra

The term "chimeric antibody" refers to an antibody in which a portion of the heavy and/or light chain is identical or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they possess the biological activity of the application (see U.S. patent No.4,816,567; and Morrison et al, proc.Natl. Acad. Sci. USA,81:6851-6855 (1984)).

"Fv" is the smallest antibody fragment that contains the complete antigen recognition and binding site. The fragment is a dimer formed by a tight non-covalent linkage of one heavy chain variable domain and one light chain variable domain. By folding of these two domains, 6 hypervariable loops (3 loops in each of the light and heavy chains) are derived, which Gao Bianhuan provides amino acid residues for the antibody to bind antigen and confers specificity to the antibody for binding to antigen. However, even a single variable domain (or half of an Fv fragment, which contains only 3 CDRs specific for an antigen) has the ability to recognize and bind antigen, although with less affinity than the complete binding site.

"Single chain Fv", also abbreviated "sFv" or "scFv", is an antibody fragment comprising V _H and V _L antibody domains linked into a single polypeptide chain. In some embodiments, the scFv polypeptide further comprises a linker polypeptide between the V _H and V _L domains that allows the scFv to form the desired structure for antigen binding. For an overview of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies,vol.113,Rosenburg and Moore eds.,Springer-Verlag,New York,pp.269-315(1994).

The term "diabody" is a small antibody fragment prepared by constructing scFv fragments (see above) using a short linker (e.g., 5-10 residues) between the V _H and V _L domains, such that the variable domains pair between the chains, rather than within the chains, to produce a bivalent fragment, i.e., a fragment having two antigen binding sites. Bispecific diabodies are heterodimers of two "crossed" scFv fragments, in which the V _H and V _L domains of the two antibodies are located on different polypeptide chains. In EP 404,097; WO 93/11161; diabodies are described fully in Hollinger et al, proc.Natl.Acad.Sci.USA,90:6444-6448 (1993).

The "humanized" form of a non-human (e.g., rodent) antibody is a chimeric antibody that includes minimal sequences from the non-human antibody. In most cases, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region (HVR) of the recipient antibody are replaced by residues from a hypervariable region of a non-human species, such as mouse, rat, rabbit or non-human mammal, having the desired antibody specificity, affinity and properties (donor antibody). In some cases, residues in the framework region of the human immunoglobulin are replaced with corresponding non-human residues. In addition, humanized antibodies may include residues that are not present in either the recipient antibody or the donor antibody. These modifications can further improve the performance of the antibody. Typically, a humanized antibody will comprise substantially all, at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are human immunoglobulin sequences. The human antibody optionally also includes at least a portion of an immunoglobulin constant region (Fc), typically a constant region of a human immunoglobulin. For specific details, reference may be made to Jones et al, nature 321:522-525 (1986); RIECHMANN ET al, nature 332:323-329 (1988); and Presta, curr.Op.struct.biol.2:593-596 (1992).

The "percent (%) amino acid sequence identity" or "homology" of the polypeptide and antibody sequences identified herein is defined as: sequence comparison is performed where conservative substitutions are considered to be part of the sequence identity, the percentage of amino acid residues in the candidate sequence that are identical to the polypeptide sequence to be compared. The percentage of amino acid sequence identity may be determined by a variety of alignment methods within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, megalign (DNASTAR), or MUSCLE software. One skilled in the art can determine suitable parameters for measuring the alignment, including any algorithms needed to achieve maximum alignment over the full length of the compared sequences. However, for purposes herein, the percent amino acid sequence identity values are generated using the sequence alignment computer program MUSCLE(Edgar,R.C.,Nucleic Acids Research 32(5):1792-1797,2004;Edgar,R.C.,BMC Bioinformatics 5(1):113,2004).

The term "Fc receptor" or "FcR" is used to describe a receptor that binds to the Fc region of an antibody. In some embodiments, the FcR of the application is one that binds an IgG antibody (a gamma receptor), including receptors of the fcyri, fcyrii, and fcyriii subclasses, including allelic variants and alternatively spliced forms of these receptors. Fcyrii receptors include fcyriia ("activating receptor") and fcyriib ("inhibiting receptor"), which have similar amino acid sequences, differing primarily in the cytoplasmic domain. The cytoplasmic domain of the activating receptor fcyriia contains an immune receptor tyrosine activation motif (ITAM). The cytoplasmic domain of the inhibition receptor fcyriib contains the Immunoreceptor Tyrosine Inhibitory Motif (ITIM) (see m.inAnnu.Rev.Immunol.15:203-234 (1997)). The term also includes allotypes, such as fcyriiia allotypes: fcgammaRIIIA-Phe 158, fcgammaRIIIA-Val 158, fcgammaRIIA-R131 and/or FcgammaRIIA-H131. At RAVETCH AND KINET, ANNU.REV.IMMUNOL 9:457-92 (1991) and Capel et al, immunomethods 4:25-34 (1994); fcRs are described in de Haas et al, J.Lab.Clin.Med.126:330-41 (1995). The term FcR in the present application encompasses other types of FcRs, including FcRs identified in the future. The term FcR also includes the neonatal receptor FcRn, which is responsible for transferring the parent IgGs to the neonate (Guyer et al, J.Immunol.117:587 (1976) and Kim et al, J.Immunol.24:249 (1994)).

The term "FcRn" refers to neonatal Fc receptor (FcRn). FcRn is structurally similar to the Major Histocompatibility Complex (MHC), consisting of non-covalent binding of the alpha chain to beta 2 microglobulin. The various functions of the neonatal Fc receptor FcRn are reviewed in GHETIE AND WARD (2000) Annu. Rev. Immunol.18,739-766. FcRn plays an important role in the passive transport of immunoglobulin IgGs from the mother to neonates and in the regulation of serum IgG levels. FcRn acts as a salvage receptor that can bind and transport endocytosed IgG in intact form within and between cells and protect them from the default degradation pathway.

The "CH1 domain" of the human IgG heavy chain constant region typically extends from amino acid 118 to amino acid 215 (EU numbering system).

The "hinge region" is generally defined as extending from Glu at position 216 to Pro at position 230 of human IgG1 (Burton, molecular immunol.22:161-206 (1985)). The hinge regions of other IgG isotypes can be aligned with the IgG1 sequence by placing the first and last cysteine residues that form the inter-heavy chain disulfide bond in the same position as IgG 1.

The "CH2 domain" of the human IgG Fc region typically extends from amino acid 231 to amino acid 340. The CH2 domain is unique in that it does not mate tightly with another region, but rather inserts two N-terminally linked branched carbohydrate chains between the two CH2 domains of the intact native IgG molecule. It is speculated that carbohydrates may serve as a surrogate for domain-to-domain pairing, helping to keep the CH2 domain stable. Burton, molecular. Immunol.22:161-206 (1985).

The "CH3" domain includes the extension from the C-terminal residue to the CH2 domain (from amino acid 341 to the C-terminal end of the antibody sequence, typically amino acid 446 or 447 of IgG) within the Fc region.

The "functional Fc fragment" has the "effector function" possessed by the native Fc region sequence. Exemplary "effector functions" include C1q binding; complement Dependent Cytotoxicity (CDC); fc receptor binding; antibody dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down-regulation of cell surface receptors (e.g., B cell receptors; BCR), and the like. Such effector functions typically require that the Fc region bind to a binding domain (e.g., an antibody variable region) and can be assessed using a variety of experimental methods well known in the art.

Antibodies to IgG Fc variants having "altered" FcR binding affinity or ADCC activity have increased or decreased FcR binding activity and/or ADCC activity compared to the parent polypeptide or polypeptide comprising the native Fc sequence. Fc variants exhibiting "enhanced binding" to FcR have a higher binding affinity (e.g., lower apparent Kd or IC ₅₀ values) to at least one FcR than the parent polypeptide or polypeptide comprising the native IgG Fc sequence. In some embodiments, the binding capacity is increased by a factor of 3, e.g., 5, 10, 25, 50, 60, 100, 150, 200, even up to a factor of 500 or the binding capacity is increased by a factor of 25% to 1000% as compared to the parent polypeptide. Fc variants exhibiting "reduced binding" to FcR have lower affinity (e.g., higher apparent Kd or IC ₅₀ values) for at least one FcR than the parent polypeptide. Its binding capacity is reduced by 40% or more compared to the parent polypeptide.

"Antibody-dependent cell-mediated cytotoxicity" or "ADCC" is a form of cytotoxicity that refers to the binding of secreted Ig to Fc receptors (FcRs) present on certain cytotoxic cells, such as natural killer cells (NK), neutrophils, and macrophages, enabling these cytotoxic effector cells to specifically bind antigen-bearing target cells, followed by killing of the target cells with cytotoxins. Antibodies "arm" cytotoxic cells and are necessary for such killing. In the major cell types mediating ADCC NK cells express fcyriii only, whereas monocytes express fcyri, fcyrii and fcyriii. FcR expression on hematopoietic cells is summarized in Table 3 at page 464 of RAVETCH AND KINET, ANNU.REV.IMMUNOL 9:457-92 (1991). The ADCC activity of the target molecule is assessed and an in vitro ADCC assay may be performed and is described in U.S. patent nos. 5,500,362 or 5,821,337. Effector cells suitable for such experiments include Peripheral Blood Mononuclear Cells (PBMC) and natural killer cells (NK). Alternatively, or in addition, ADCC activity of the target molecule may also be assessed in vivo, for example as described in an animal model as disclosed in Clynes et al PNAS (USA) 95:652-656 (1998).

Polypeptides comprising an Fc region variant that are experimentally substantially the same in number as wild-type IgG Fc polypeptides (or parent polypeptides) are more effective in mediating ADCC in vitro or in vivo when they exhibit "enhanced ADCC activity" or are capable of mediating ADCC effects more effectively in the presence of human effector cells than wild-type IgG Fc polypeptides or parent polypeptides. Such variants are typically identified using any in vitro ADCC assay known in the art, such as assays or methods for identifying ADCC activity, e.g., in animal models, etc. In some embodiments, such variants mediate ADCC with a 5 to 100 fold, e.g., 25 to 50 fold increase in efficiency compared to the wild-type Fc (or parent polypeptide).

"Complement dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by binding of the first component of the complement system (C1 q) to antibodies (subclasses of appropriate structure) that bind to cognate antigens. To assess complement activation, CDC experiments can be performed as described in Gazzano-Santoro et al, J.Immunol. Methods 202:163 (1996). Polypeptide variants having altered amino acid sequences of the Fc region and increased or decreased C1q binding capacity are described in U.S. Pat. No.6,194,551B1 and WO 99/51642. The contents of these patent publications are expressly incorporated herein by reference. See also Idusogie et al J.Immunol.164:4178-4184 (2000).

Unless otherwise indicated, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate versions of each other and encode the same amino acid sequence. The nucleotide sequence encoding a protein or RNA may also include introns, e.g., the nucleotide sequence encoding a protein may in some forms comprise introns.

The term "operably linked" refers to a functional linkage between a regulatory sequence and a heterologous nucleotide sequence such that the latter is expressed. For example, a first nucleotide sequence is operably linked to a second nucleotide sequence when the first nucleotide sequence is in a functional relationship with the second nucleotide sequence. For example, a promoter is operably linked to a coding sequence if it affects the transcription or expression of the coding sequence. Typically, operably linked DNA sequences are contiguous and, if necessary, two protein coding regions can be linked in the same reading frame.

"Homology" refers to sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. If the same position of two compared sequences is the same base or amino acid monomer subunit, for example, the same position of both DNA molecules is adenine, then both DNA molecules are homologous at that position. The percentage of homology between two sequences refers to the function of the ratio of the number of matching or homologous positions to the total number of positions shared by the two sequences multiplied by 100. For example, if 6 of the 10 positions in two sequences are matched or homologous, the two sequences are 60% homologous. For example, the DNA sequences ATTGCC and TATGGC have 50% homology. In general, when two sequences are aligned, alignment is performed with the aim of obtaining maximum homology.

An "effective amount" of an anti-TRAIL antibody or composition disclosed herein refers to an amount sufficient to achieve a particular purpose. The "effective amount" may be determined empirically and by methods known in connection with the purpose.

The term "therapeutically effective amount" refers to an amount of an anti-TRAIL antibody or composition thereof disclosed herein that is effective to treat a disease or condition in an individual, i.e., an amount sufficient to reduce or ameliorate the severity and/or duration of the disease or one or more symptoms thereof; preventing disease progression, causing regression of the condition, preventing recurrence, development, onset, or progression of one or more symptoms associated with the disease, detecting the disease, or enhancing/improving the prophylactic or therapeutic effect of another therapy (e.g., a prophylactic or therapeutic agent). For example, in the case of cancer, a therapeutically effective amount of an anti-TRAIL antibody or composition thereof is one that reduces the number of cancer cells; reducing the size or weight of the tumor; inhibit (i.e., slow or preferably stop to some extent) infiltration of peripheral organs by tumor cells; inhibit (i.e., slow or preferably stop to some extent) tumor metastasis; inhibit to some extent the growth of tumors, and/or alleviate to some extent one or more symptoms associated with cancer. The anti-TRAIL antibodies or compositions thereof disclosed herein are capable of preventing or inhibiting to some extent the binding of TRAIL to its receptor DR4/DR5, except for an apoptosis signaling pathway, a TRAIL-mediated non-apoptotic signal (such as NF-kB, MAPK or Akt) that promotes development, survival and proliferation. All of these activities may be inhibited by the antibodies described herein.

As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" refers to materials that are not biologically active or otherwise undesirable, e.g., that can be added to a pharmaceutical composition administered to a patient without causing significant adverse biological reactions or interacting in a deleterious manner with any of the other components of the composition in which they are contained. The pharmaceutically acceptable carrier or excipient preferably meets the desired criteria for toxicology or manufacturing testing and/or is contained in inactive ingredient guidelines established by the U.S. food and drug administration.

The embodiments of the application described herein should be understood to include embodiments consisting of … … and/or consisting essentially of … ….

Reference herein to "about" is a numerical value or parameter, including (and describing) variations on the value or parameter itself. For example, a description relating to "about X" includes a description of "X".

As used herein, reference to a value or parameter that is "not (not)" generally means and describes "other than (other than)" a value or parameter. For example, the method cannot be used to treat type X cancers, meaning that the method is generally used to treat other types of cancers in addition to type X cancers.

As used herein and in the claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

Anti-TRAIL antibodies

In one aspect, the application provides anti-TRAIL antibodies that specifically bind TRAIL. Such anti-TRAIL antibodies include, but are not limited to, humanized antibodies, chimeric antibodies, mouse antibodies, human antibodies, and antibody molecules comprising heavy and/or light chain CDRs as described herein. In one aspect, the application provides isolated antibodies that bind to TRAIL. Contemplated anti-TRAIL antibodies include, for example, full length anti-TRAIL antibodies (e.g., full length IgG1 or IgG 4), anti-TRAIL single chain antibodies, anti-TRAIL Fc fusion proteins, multi-specific (e.g., bispecific) anti-TRAIL antibodies, anti-TRAIL immunoconjugates, and the like. In some embodiments, the anti-TRAIL antibody is a full length antibody (e.g., full length IgG1 or IgG 4) or an antigen-binding fragment thereof, which specifically binds TRAIL. In some embodiments, the anti-TRAIL antibody is a Fab, fab ', F (ab) ' ₂, fab ' -SH, single chain Fv (scFv), fv fragment, dAb, fd, nanobody (nanobody), diabody (diabody), or linear antibody. In some embodiments, an antibody that specifically binds TRAIL refers to an antibody that binds TRAIL with at least 10-fold or more (including, for example, 10 ²、10³、10⁴、10⁵、10⁶, or 10 ⁷ -fold) greater affinity than non-target binding affinity. In some embodiments, non-target refers to an antigen that is not TRAIL. Binding affinity can be determined by methods known in the art, such as ELISA, fluorescence Activated Cell Sorting (FACS) analysis, or Radioimmunoassay (RIA). Kd values can be determined by methods known in the art, such as Surface Plasmon Resonance (SPR) techniques or Biological Layer Interferometry (BLI).

While anti-TRAIL antibodies comprising human sequences (e.g., human heavy and light chain variable domains comprising human CDR sequences) are discussed broadly herein, non-human anti-TRAIL antibodies are also contemplated. In some embodiments, the non-human anti-TRAIL antibodies include human CDR sequences and non-human framework region sequences of anti-TRAIL antibodies described herein, and in some embodiments, the non-human framework region sequences include any sequences for producing heavy and/or light chain variable domains using one or more human CDR sequences as described herein, including, for example, mammals, such as mice, rats, rabbits, pigs, cattle (e.g., cattle, bulls, buffalo), deer, sheep, goats, chickens, cats, dogs, ferrets, primates (e.g., apes, macaques), and the like. In some embodiments, the non-human anti-TRAIL antibodies include anti-TRAIL antibodies produced by grafting one or more human CDR sequences described herein into a non-human framework region (e.g., a murine or chicken framework region sequence).

An exemplary human TRAIL comprises the complete amino acid sequence of SEQ ID NO:93 or an amino acid sequence set forth in SEQ ID NO: 93.

In some embodiments, the anti-TRAIL antibodies described herein specifically recognize an epitope in human TRAIL. In some embodiments, the anti-TRAIL antibody cross-reacts with TRAIL of other species than human. In some embodiments, the anti-TRAIL antibody is fully specific for human TRAIL and does not cross-react with other non-human species.

In some embodiments, the anti-TRAIL antibody cross-reacts with at least one allelic variant of TRAIL protein (or fragment thereof). In some embodiments, the allelic variant has up to 30 (e.g., 1,2, 3, 4, 5,6, 7, 8, 9,10, 15, 20, 25, or 30) amino acid substitutions (e.g., conservative substitutions) as compared to the naturally occurring TRAIL protein (or fragment thereof). In some embodiments, the anti-TRAIL antibody does not cross-react with any allelic variants of TRAIL protein (or fragment thereof).

In some embodiments, the anti-TRAIL antibody cross-reacts with at least one intervarietal variant of TRAIL protein. In some embodiments, for example, the TRAIL protein (or fragment thereof) is human TRAIL, and the intervarietal variant of the TRAIL protein (or fragment thereof) is a variant in cynomolgus monkey. In some embodiments, the anti-TRAIL antibody does not cross-react with any inter-variant TRAIL protein.

In some embodiments, any of the anti-TRAIL antibodies as described herein, the anti-TRAIL antibody comprises an antibody heavy chain constant region and an antibody light chain constant region. In some embodiments, the anti-TRAIL antibody comprises an IgG1 type heavy chain constant region. In some embodiments, the anti-TRAIL antibody comprises an IgG2 type heavy chain constant region. In some embodiments, the anti-TRAIL antibody comprises an IgG3 type heavy chain constant region. In some embodiments, the anti-TRAIL antibody comprises an IgG4 type heavy chain constant region. In some embodiments, the heavy chain constant region comprises (including consisting of … or consisting essentially of …) the amino acid sequence SEQ ID NO 89. In some embodiments, the heavy chain constant region comprises (including consisting of … or consisting essentially of …) the amino acid sequence SEQ ID NO 90. In some embodiments, the anti-TRAIL antibody comprises a kappa light chain constant region. In some embodiments, the light chain constant region comprises (including consisting of … or consisting essentially of …) the amino acid sequence SEQ ID NO 91. In some embodiments, the anti-TRAIL antibody comprises a lambda light chain constant region. In some embodiments, the light chain constant region comprises (including consisting of … or consisting essentially of …) the amino acid sequence SEQ ID NO 92. In some embodiments, the anti-TRAIL antibody comprises an antibody heavy chain variable domain and an antibody light chain variable domain.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, HC-CDR3 comprising the amino acid sequence

18, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 29, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:18; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 29, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 50; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 70.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 50 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 50; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 70, or a variant thereof, having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 70. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence of SEQ ID No. 50, and V _L, the V _L comprises the amino acid sequence of SEQ ID No. 70.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, HC-CDR3 comprising the amino acid sequence

18, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, LC-CDR2 comprising the amino acid sequence SEQ ID NO:37, LC-CDR3 comprising the amino acid sequence SEQ ID NO:41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, LC-CDR2 comprising the amino acid sequence SEQ ID NO:37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:41.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 51; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 71.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 51 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 51; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 71 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 71. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID NO:51, and V _L, the V _L comprises the amino acid sequence SEQ ID NO:71.

18, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 31, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 31, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 52; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 72.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 52 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 52; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 72 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 72. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID NO:52, and V _L, the V _L comprises the amino acid sequence SEQ ID NO:72.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, HC-CDR3 comprising the amino acid sequence

19, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:19; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 53; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 73.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 53 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 53; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 73 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 73. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence of SEQ ID No. 53, and V _L, the V _L comprises the amino acid sequence of SEQ ID No. 73.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, HC-CDR3 comprising the amino acid sequence

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 19; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 54; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 74.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 54 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 54; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 74 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 74. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID NO:54, and V _L, the V _L comprises the amino acid sequence SEQ ID NO:74.

20, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:20; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 55; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 as set forth in amino acid sequence SEQ ID NO 75 as V _L.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 55 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 55; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 75 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 75. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence of SEQ ID No. 55, and V _L, the V _L comprises the amino acid sequence of SEQ ID No. 75.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, HC-CDR3 comprising the amino acid sequence

21, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 21; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 56; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 76.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 56 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 56; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 76, or a variant thereof, having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 76. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID NO:56, and V _L, the V _L comprises the amino acid sequence SEQ ID NO:76.

22, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 22; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 57; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 74.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 57 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 57; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 74 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 74. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 57, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 74.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 33, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 39, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 43, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 33, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 39, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 43.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 58; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 77.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 58 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 58; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 77 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 77. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID NO:58, and V _L, the V _L comprises the amino acid sequence SEQ ID NO:77.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO.7, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, HC-CDR3 comprising the amino acid sequence

24, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO 34, LC-CDR2 comprising the amino acid sequence SEQ ID NO 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 7, HC-CDR2 comprising the amino acid sequence SEQ ID NO.14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO 34, LC-CDR2 comprising the amino acid sequence SEQ ID NO 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO 44.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 59; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 78.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 59 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 59; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 78 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 78. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 59, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 78.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, HC-CDR3 comprising the amino acid sequence

24, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO.14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 60; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 79.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 60 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 60; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 79 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 79. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 60, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 79.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 61; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 80.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 61 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 61; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 80 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 80. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 61, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 80.

24, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, LC-CDR3 comprising the amino acid sequence SEQ ID NO:45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO.14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 62; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 81.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 62 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 62; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 81 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 81. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence of SEQ ID No. 62, and V _L, the V _L comprises the amino acid sequence of SEQ ID No. 81.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, HC-CDR3 comprising the amino acid sequence

24, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:35, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, LC-CDR3 comprising the amino acid sequence SEQ ID NO:45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:35, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:45.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO. 63; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 82.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 63 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 63; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 82 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 82. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 63, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 82.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, HC-CDR3 comprising the amino acid sequence

25, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, LC-CDR3 comprising the amino acid sequence SEQ ID NO:45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 25; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 64; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 83.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 64 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 64; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 83 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 83. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 64, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 83.

25, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 46, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 25; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 46.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 65; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 84.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 65 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 65; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 84 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 84. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 65, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 84.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, HC-CDR3 comprising the amino acid sequence SEQ ID NO 26, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:36, LC-CDR2 comprising the amino acid sequence SEQ ID NO:40, LC-CDR3 comprising the amino acid sequence SEQ ID NO:47, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 26; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:36, LC-CDR2 comprising the amino acid sequence SEQ ID NO:40, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:47.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 66; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 85.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 66 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 66; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 85 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 85. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 66, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 85.

24, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, LC-CDR3 comprising the amino acid sequence SEQ ID NO:48, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO.14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:48.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 67; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 86.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 67 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 67; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 86 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 86. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 67, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 86.

27, Or a variant of said V _H, comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 27; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO: 68; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 87.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 68 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 68; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 87 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 87. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 68, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 87.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, LC-CDR3 comprising the amino acid sequence SEQ ID NO:49, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:49.

In some embodiments, the anti-TRAIL antibody comprises V _H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in V _H of amino acid sequence SEQ ID NO 69; and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 88.

In some embodiments, the anti-TRAIL antibody comprises: v _H, said V _H comprising the amino acid sequence SEQ ID No. 69 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID No. 69; and V _L, the V _L comprising the amino acid sequence SEQ ID No. 88 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence SEQ ID No. 88. In some embodiments, the anti-TRAIL antibody comprises V _H, the V _H comprises the amino acid sequence SEQ ID No. 69, and V _L, the V _L comprises the amino acid sequence SEQ ID No. 88.

In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in table 4 herein. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in table 4 herein.

In some embodiments, the functional epitope can be resolved by combining alanine scanning methods. In this process, combinatorial alanine scanning techniques can be used to identify amino acids in TRAIL proteins that are necessary for interaction with anti-TRAIL antibodies. In some embodiments, the epitope is conformational, and the crystal structure of an anti-TRAIL antibody that binds to TRAIL protein may be employed to identify the epitope.

In some embodiments, the application provides antibodies that competitively bind TRAIL with any of the anti-TRAIL antibodies described herein. In some embodiments, antibodies are provided that are capable of competitively binding to an epitope on TRAIL with any of the anti-TRAIL antibodies described herein. In some embodiments, anti-TRAIL antibodies are provided that bind to the same epitope as an anti-TRAIL antibody molecule comprising V _H and V _L, wherein the V _H comprises the amino acid sequence set forth in any one of SEQ ID NOs:50-69, and the V _L comprises the amino acid sequence set forth in any one of SEQ ID NOs: 70-88. In some embodiments, anti-TRAIL antibodies are provided that competitively bind to TRAIL with anti-TRAIL antibodies comprising V _H and V _L, wherein the V _H comprises the amino acid sequence set forth in any one of SEQ ID NOs:50-69, and the V _L comprises the amino acid sequence set forth in any one of SEQ ID NOs: 70-88.

In some embodiments, competition experiments can be used to identify monoclonal antibodies that competitively bind TRAIL with the anti-TRAIL antibodies described herein. Competition experiments can determine whether two antibodies bind to the same epitope by recognizing the same or spatially overlapping epitopes or by one antibody competitively inhibiting the binding of the other antibody to the antigen. In certain embodiments, such a competing antibody binds to the same epitope as the antibodies described herein. Some exemplary competition experiments include, but are not limited to, routine experiments as mentioned in Harlow and Lane(1988)Antibodies:A Laboratory Manual ch.14(Cold Spring Harbor Laboratory,Cold Spring Harbor,N.Y.). A detailed exemplary method for resolving epitopes to which antibodies bind is described in Morris(1996)"Epitope Mapping Protocols,"in Methods in Molecular Biology vol.66(Humana Press,Totowa,N.J.). In some embodiments, each antibody is said to bind to the same epitope if it blocks 50% or more of the binding of the other antibody. In some embodiments, the antibody that competes with an anti-TRAIL antibody described herein is a chimeric, humanized, or fully human antibody.

Exemplary anti-TRAIL antibody sequences are shown in tables 2,3, wherein CDR numbering is performed according to Chothia definition. Those skilled in the art will recognize that there are a variety of known algorithms to predict CDR positions and define antibody light and heavy chain variable regions. Antibodies comprising CDRs, V _H, and/or V _L sequences of antibodies as described herein, but based on predictive algorithms other than those exemplified in the tables below, are also within the scope of the application.

TABLE 2 exemplary anti-TRAIL antibody CDR sequences

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Table 3 exemplary sequences

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Full-length anti-TRAIL antibodies

In some embodiments, the anti-TRAIL antibody is a full-length anti-TRAIL antibody. In some embodiments, the full-length anti-TRAIL antibody is IgA, igD, igE, igG or IgM. In some embodiments, the full length anti-TRAIL antibody comprises an IgG constant region, e.g., a constant region of IgG1, igG2, igG3, igG4, or a variant thereof. In some embodiments, the full length anti-TRAIL antibody comprises a lambda light chain constant region. In some embodiments, the full length anti-TRAIL antibody comprises a kappa light chain constant region. In some embodiments, the full length anti-TRAIL antibody is a full length human anti-TRAIL antibody. In some embodiments, the full-length anti-TRAIL antibody comprises a mouse immunoglobulin Fc sequence. In some embodiments, the full-length anti-TRAIL antibody comprises an Fc sequence that has been altered or otherwise altered such that it has enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) effector functions.

Thus, for example, in some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided that specifically binds TRAIL. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG2 constant region is provided that specifically binds TRAIL. In some embodiments, the IgG2 is human IgG2. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG3 constant region is provided that specifically binds TRAIL. In some embodiments, the IgG3 is human IgG3. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided that specifically binds TRAIL. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 1-11 or a variant thereof comprising up to about 3 (e.g., 1, 2 or 3) amino acid substitutions;

HC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 12-17 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions; and HC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 18-28 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions; and b) a light chain variable domain comprising: LC-CDR1 comprising an amino acid sequence as set forth in any one of SEQ ID NOs 29-36 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions, LC-CDR2 comprising an amino acid sequence as set forth in any one of SEQ ID NOs 37-40 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions; and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 41-49 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions. In some embodiments, the IgG1 is human

IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG2 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 1-11 or a variant thereof comprising up to about 3 (e.g., 1, 2 or 3) amino acid substitutions;

HC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 12-17 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions; and HC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 18-28 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 29-36 or a variant thereof comprising up to about 3 (e.g. 1,2 or 3) amino acid substitutions; LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 37-40 or a variant thereof comprising up to about 3 (e.g. 1,2 or 3) amino acid substitutions; and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 41-49 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions. In some embodiments, the IgG2 is human

IgG2. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG3 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence set forth in any one of SEQ ID NOs 1-11 or a variant thereof comprising up to about 3 (e.g., 1, 2 or 3) amino acid substitutions, HC-CDR2 comprising the amino acid sequence set forth in any one of SEQ ID NOs 12-17 or a variant thereof comprising up to about 3 (e.g., 1, 2 or 3) amino acid substitutions; and HC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 18-28 or a variant thereof comprising up to about 3 (e.g., 1, 2 or 3) amino acid substitutions; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 29-36 or a variant thereof comprising up to about 3 (e.g. 1, 2 or 3) amino acid substitutions; LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 37-40 or a variant thereof comprising up to about 3 (e.g. 1, 2 or 3) amino acid substitutions; and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 41-49 or a variant thereof comprising up to about 3 (e.g., 1, 2 or 3) amino acid substitutions. In some embodiments, the IgG3 is human

IgG3. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 1-11 or a variant thereof comprising up to about 3 (e.g., 1, 2 or 3) amino acid substitutions;

HC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 12-17 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions; and HC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 18-28 or a variant thereof comprising up to about 3 (e.g., 1,2 or 3) amino acid substitutions; and b) a light chain variable domain comprising: LC-CDR1 comprising an amino acid sequence as set forth in any one of SEQ ID NOs 29-36 or a variant thereof comprising up to about 3 (e.g. 1,2 or 3) amino acid substitutions, LC-CDR2 comprising an amino acid sequence as set forth in any one of SEQ ID NOs 37-40 or a variant thereof comprising up to about 3 (e.g. 1,2 or 3) amino acid substitutions, and LC-CDR3 comprising an amino acid sequence as set forth in any one of SEQ ID NOs 41-49 or a variant thereof comprising up to about 3 (e.g. 1,2 or 3) amino acid substitutions. In some embodiments, the IgG4 is human

IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence set forth in any one of SEQ ID NOs 1-11, HC-CDR2 comprising the amino acid sequence set forth in any one of SEQ ID NOs 12-17, and HC-CDR3 comprising the amino acid sequence set forth in any one of SEQ ID NOs 18-28, or a variant of said heavy chain variable domain comprising up to about 5 (e.g., 1,2,3,4 or 5) amino acid substitutions in the HC-CDR sequence; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 29-36, LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 37-40, and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 41-49, or a variant of said light chain variable domain comprising up to about 5 (e.g. 1,2,3,4 or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises a) a heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence set forth in any one of SEQ ID NOs 1-11, HC-CDR2 comprising the amino acid sequence set forth in any one of SEQ ID NOs 12-17, and HC-CDR3 comprising the amino acid sequence set forth in any one of SEQ ID NOs 18-28, or a variant of said heavy chain variable domain comprising up to about 5 (e.g., 1,2,3,4 or 5) amino acid substitutions in the HC-CDR sequence; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 29 to 36, LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 37 to 40, and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 41 to 49, or a variant of said light chain variable domain comprising up to about 5 (e.g. 1,2,3,4 or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 1-11, HC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 12-17, and HC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 18-28; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 29-36, LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 37-40, and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 41-49. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 1-11, HC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 12-17, and HC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 18-28; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs 29-36, LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs 37-40, and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs 41-49. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:18; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 29, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, LC-CDR2 comprising the amino acid sequence SEQ ID NO:37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:41. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 31, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:19; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 19; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:20; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 21; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 22; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 33, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 39, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 43. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 7, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO 34, LC-CDR2 comprising the amino acid sequence SEQ ID NO 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO 44. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:35, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:45. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 25; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 25; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 46. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 26; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:36, LC-CDR2 comprising the amino acid sequence SEQ ID NO:40, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:47. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:48. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 27; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:49. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:18; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 29, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, LC-CDR2 comprising the amino acid sequence SEQ ID NO:37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:41. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 31, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:19; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 19; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:20; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 21; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 22; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:42. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 33, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 39, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 43. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 7, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO 34, LC-CDR2 comprising the amino acid sequence SEQ ID NO 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO 44. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:35, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:45. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 25; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 25; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 46. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 26; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:36, LC-CDR2 comprising the amino acid sequence SEQ ID NO:40, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:47. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:48. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 27; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a) A heavy chain variable domain comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:49. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: a heavy chain variable domain (V _H), said V _H comprising an amino acid sequence as set forth in any one of SEQ ID NOs 50-69 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to an amino acid sequence as set forth in any one of SEQ ID NOs 50-69; and a light chain variable domain (V _L), the V _L comprising an amino acid sequence set forth in any one of SEQ ID NOs 70-88 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence set forth in any one of SEQ ID NOs 70-88. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG2 constant region is provided, wherein the anti-TRAIL antibody comprises: a heavy chain variable domain (V _H), said V _H comprising an amino acid sequence as set forth in any one of SEQ ID NOs 50-69 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to an amino acid sequence as set forth in any one of SEQ ID NOs 50-69; and a light chain variable domain (V _L), the V _L comprising an amino acid sequence set forth in any one of SEQ ID NOs 70-88 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence set forth in any one of SEQ ID NOs 70-88. In some embodiments, the IgG2 is human IgG2. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG3 constant region is provided, wherein the anti-TRAIL antibody comprises: a heavy chain variable domain (V _H), said V _H comprising an amino acid sequence as set forth in any one of SEQ ID NOs 50-69 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to an amino acid sequence as set forth in any one of SEQ ID NOs 50-69; and a light chain variable domain (V _L), the V _L comprising an amino acid sequence set forth in any one of SEQ ID NOs 70-88 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence set forth in any one of SEQ ID NOs 70-88. In some embodiments, the IgG3 is human IgG3. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: a heavy chain variable domain (V _H), said V _H comprising an amino acid sequence as set forth in any one of SEQ ID NOs 50-69 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to an amino acid sequence as set forth in any one of SEQ ID NOs 50-69; and a light chain variable domain (V _L), the V _L comprising an amino acid sequence set forth in any one of SEQ ID NOs 70-88 or a variant thereof having at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence set forth in any one of SEQ ID NOs 70-88. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: heavy chain variable domain (V _H), the V _H comprising the amino acid sequence set forth in any one of SEQ ID NOs:50-69, and light chain variable domain (V _L), the V _L comprising the amino acid sequence set forth in any one of SEQ ID NOs: 70-88. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of an amino acid sequence

SEQ ID NO. 89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: heavy chain variable domain (V _H), the V _H comprising the amino acid sequence set forth in any one of SEQ ID NOs:50-69, and light chain variable domain (V _L), the V _L comprising the amino acid sequence set forth in any one of SEQ ID NOs: 70-88. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of an amino acid sequence

SEQ ID NO. 90 and the light chain constant region comprise or consist of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 50 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 50; and V _L comprising the amino acid sequence SEQ ID NO. 70 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 70. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 51 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 51; and V _L comprising the amino acid sequence SEQ ID NO. 71 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 71. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 52 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 52; and V _L comprising the amino acid sequence SEQ ID NO. 72 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 72. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 53 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 53; and V _L comprising the amino acid sequence SEQ ID NO. 73 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 73. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 54 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 54; and V _L comprising the amino acid sequence SEQ ID NO. 74 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 74. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 55 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 55; and V _L comprising the amino acid sequence SEQ ID NO. 75 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 75. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 56 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 56; and V _L comprising the amino acid sequence SEQ ID NO. 76 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 76. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 57 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 57; and V _L comprising the amino acid sequence SEQ ID NO. 74 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 74. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 58 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 58; and V _L comprising the amino acid sequence SEQ ID NO. 77 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 77. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 59 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 59; and V _L comprising the amino acid sequence SEQ ID NO. 78 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 78. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 60 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 60; and V _L comprising the amino acid sequence SEQ ID NO. 79 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 79. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 61 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 61; and V _L comprising the amino acid sequence SEQ ID NO. 80 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 80. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 62 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 62; and V _L comprising the amino acid sequence SEQ ID NO. 81 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 81. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 63 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 63; and V _L comprising the amino acid sequence SEQ ID NO. 82 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 82. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 64 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 64; and V _L comprising the amino acid sequence SEQ ID NO. 83 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 83. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 65 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 65; and V _L comprising the amino acid sequence SEQ ID NO. 84 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 84. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 66 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 66; and V _L comprising the amino acid sequence SEQ ID NO. 85 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 85. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 67 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 67; and V _L comprising the amino acid sequence SEQ ID NO. 86 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 86. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 68 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 68; and V _L comprising the amino acid sequence SEQ ID NO. 87 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 87. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG1 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 69 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 69; and V _L comprising the amino acid sequence SEQ ID NO. 88 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 88. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:89 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 50 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 50; and V _L comprising the amino acid sequence SEQ ID NO. 70 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 70. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 51 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 51; and V _L comprising the amino acid sequence SEQ ID NO. 71 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 71. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 52 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 52; and V _L comprising the amino acid sequence SEQ ID NO. 72 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 72. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 53 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 53; and V _L comprising the amino acid sequence SEQ ID NO. 73 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 73. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 54 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 54; and V _L comprising the amino acid sequence SEQ ID NO. 74 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 74. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 55 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 55; and V _L comprising the amino acid sequence SEQ ID NO. 75 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 75. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 56 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 56; and V _L comprising the amino acid sequence SEQ ID NO. 76 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 76. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 57 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 57; and V _L comprising the amino acid sequence SEQ ID NO. 74 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 74. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 58 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 58; and V _L comprising the amino acid sequence SEQ ID NO. 77 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 77. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 59 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 59; and V _L comprising the amino acid sequence SEQ ID NO. 78 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 78. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 60 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 60; and V _L comprising the amino acid sequence SEQ ID NO. 79 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 79. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 61 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 61; and V _L comprising the amino acid sequence SEQ ID NO. 80 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 80. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 62 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 62; and V _L comprising the amino acid sequence SEQ ID NO. 81 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 81. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 63 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 63; and V _L comprising the amino acid sequence SEQ ID NO. 82 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 82. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 64 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 64; and V _L comprising the amino acid sequence SEQ ID NO. 83 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 83. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 65 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 65; and V _L comprising the amino acid sequence SEQ ID NO. 84 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 84. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 66 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 66; and V _L comprising the amino acid sequence SEQ ID NO. 85 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 85. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 67 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 67; and V _L comprising the amino acid sequence SEQ ID NO. 86 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 86. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 68 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 68; and V _L comprising the amino acid sequence SEQ ID NO. 87 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 87. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

In some embodiments, a full length anti-TRAIL antibody comprising an IgG4 constant region is provided, wherein the anti-TRAIL antibody comprises: v _H comprising the amino acid sequence SEQ ID No. 69 or a variant thereof having at least about 80% sequence identity to the amino acid sequence SEQ ID No. 69; and V _L comprising the amino acid sequence SEQ ID NO. 88 or a variant thereof, said variant having at least about 80% sequence identity to the amino acid sequence SEQ ID NO. 88. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

Binding affinity

Binding affinity is expressed in Kd, koff, kon or Ka. As used herein, the term Koff refers to the rate constant of dissociation of an antibody from an antigen/antibody complex, as determined by a kinetic selection device. The term Kon refers to the binding rate constant of an antibody to an antigen to form an antigen/antibody complex. The equilibrium dissociation constant Kd as used herein refers to the dissociation constant at which a particular antibody antigen interacts, meaning that in a solution of an antibody molecule, the antigen occupies half of all antibody binding sites and the concentration of antigen required to reach equilibrium is equal to Koff/Kon. Determination of Kd assumes that all binding molecules are in solution. In the case of antibody attachment to the cell wall, for example in yeast expression systems, the corresponding equilibrium dissociation rate constant is expressed in EC ₅₀, which is a good approximation of Kd. The affinity binding constant Ka is the inverse of the dissociation constant Kd.

The dissociation constant (Kd) can be used as an indicator of the affinity of the reactive antibody moiety for the antigen. For example, the interactions between biomolecules can be analyzed by Scatchard method using antibodies labeled with various markers, and Biacore instrument (manufactured by Amersham Biosciences) according to user manual or attached kit, by surface plasmon resonance. The Kd values obtained using these methods are expressed in units M. Antibodies that specifically bind to a target may have, for example, kd values of 10 ^-7M、≤10^-8M、≤10^-9M、≤10^-10M、≤10^-11M、≤10^-12 M or 10 ^-13 M.

The binding specificity of an antibody can be determined experimentally by methods known in the art. These methods include, but are not limited to, western blots, ELISA-, RIA-, ECL-, IRMA-, EIA-, BIAcore assays, peptide scans, and the like.

In some embodiments, the anti-TRAIL antibody specifically binds to TRAIL targets with a Kd value of 10 ^-7 M to 10 ^-13 M (e.g., 10 ^-7 M to 10 ^-13M、10^-8 M to 10 ^-13M、10^-9 M to 10 ^-13 M or 10 ^-10 M to 10 ^-12 M). Thus, in some embodiments, the Kd value of binding between an anti-TRAIL antibody and TRAIL is from 10 ^-7 M to 10 ^-13M、1×10^-7 M to 5 x 10 ^-13M、10^-7 M to 10 ^-12M、10^-7 M to 10 ^-11M、10^-7 M to 10 ^-10M、10^-7 M to 10 ^-9M、10^-8 M to 10 ^-13M、1×10^-8 M to 5 x 10 ^-13M、10^-8 M to 10 ^- ¹²M、10^-8 M to 10 ^-11M、10^-8 M to 10 ^-10M、10^-8 M to 10 ^-9M、5×10^-9 M to 1x 10 ^-13M、5×10^-9 M to 1x 10 ^-12M、5×10^-9 M to 1x 10 ^-11M、5×10^-9 M to 1x 10 ^-10M、10^-9 M to 10 ^-13M、10^-9 M to 10 ^-12M、10^-9 M to 10 ^-11M、10^-9 M to 10 ^-10M、5×10^-10 M to 1x 10 ^-13M、5×10^-10 M to 1x 10 ^-12M、5×10^-10 M to 1x 10 ^-11M、10^-10 M to 10 ^- ¹³M、1×10^-10 M to 5 x 10 ^-13M、1×10^-10 M to 1x 10 ^-12M、1×10^-10 M to 5 x 10 ^-12M、1×10^-10 M to 1x 10 ^- ¹¹M、10^-11 M to 10 ^-13M、1×10^-11 M to 5 x 10 ^-13M、10^-11 M to 10 ^-12M、10^-12 M to 10 ^-13 M. In some embodiments, the Kd value of binding between an anti-TRAIL antibody and TRAIL is 10 ^-7 M to 10 ^-13 M.

In some embodiments, the Kd value of binding between the anti-TRAIL antibody and the non-target is higher than the Kd value of the anti-TRAIL antibody and the target, and in some embodiments cited herein, the binding affinity of the anti-TRAIL antibody to the target (e.g., TRAIL) is higher than the binding affinity of the anti-TRAIL antibody to the non-target. In some embodiments, non-target refers to an antigen other than TRAIL. In some embodiments, the anti-TRAIL antibodies (directed against TRAIL) have a difference in Kd values between the binding of the anti-TRAIL antibodies to the non-TRAIL targets of at least 10-fold, e.g., 10-100-fold, 100-1000-fold, 10 ³-10⁴ -fold, 10 ⁴-10⁵ -fold, 10 ⁵-10⁶ -fold, 10 ⁶-10⁷ -fold, 10 ⁷-10⁸ -fold, 10 ⁸-10⁹ -fold, 10 ⁹ -fold

10 ¹⁰ Times, 10 ¹⁰-10¹¹ times, 10 ¹¹-10¹² times.

In some embodiments, the anti-TRAIL antibody binds to a non-target with a Kd value of 10 ^-1 M to 10 ^-6 M (e.g., 10 ^- ¹ M to 10 ^-6M、10^-1 M to 10 ^-5M、10^-2 M to 10 ^-4 M). In some embodiments, the non-target refers to an antigen other than TRAIL. Thus, in some embodiments, the Kd value of binding between an anti-TRAIL antibody and a non-TRAIL target is 10 ^-1 M to 10 ^-6M、1×10^-1 M to 5 x 10 ^-6M、10^-1 M to 10 ^-5M、1×10^-1 M to 5 x 10 ^-5M、10^-1 M to 10 ^-4M、1×10^-1 M to 5 x 10 ^-4M、10^-1 M to 10 ^-3M、1×10^-1 M to 5 x 10 ^-3M、10^-1 M to 10 ^-2M、10^-2 M to 10 ^-6M、1×10^-2 M to 5 x 10 ^-6M、10^-2 M to 10 ^- ⁵M、1×10^-2 M to 5 x 10 ^-5M、10^-2 M to 10 ^-4M、1×10^-2 M to 5 x 10 ^-4M、10^-2 M to 10 ^-3M、10^-3 M to 10 ^-6M、1×10^-3 M to 5 x 10 ^-6M、10^-3 M to 10 ^-5M、1×10^-3 M to 5 x 10 ^-5M、10^-3 M to 10 ^-4M、10^-4 M to 10 ^-6M、1×10^-4 M to 5 x 10 ^-6M、10^-4 M to 10 ^-5M、10^-5 M to 10 ^-6 M.

In some embodiments, when referring to an anti-TRAIL antibody specifically recognizing TRAIL targets with high binding affinity and binding non-targets with low binding affinity, the anti-TRAIL antibody binds to TRAIL targets with a Kd value of 10 ^-7 M to 10 ^- ¹³ M (e.g., 10 ^-7 M to 10 ^-13M、10^-8 M to 10 ^-13M、10^-9 M to 10 ^-13M、10^-10 M to 10 ^-12 M) and with a Kd value of 10 ^-1 M to 10 ^-6 M (e.g., 10 ^-1 M to 10 ^-6M、10^-1 M to 10 ^-5M、10^-2 M to 10 ^-4 M) with non-targets.

In some embodiments, when referring to an anti-TRAIL antibody specifically recognizing TRAIL, the binding affinity of the anti-TRAIL antibody is compared to the binding affinity of a control anti-TRAIL antibody (e.g., MAB 375-500). In some embodiments, the Kd value for binding between a control anti-TRAIL antibody and TRAIL may be at least 2-fold, e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-100-fold, 100-1000-fold, 10- ³-10⁴ -fold, of the Kd value for binding between an anti-TRAIL antibody and TRAIL described herein.

Nucleic acid

Nucleic acid molecules encoding anti-TRAIL antibodies are also contemplated. In some embodiments, a nucleic acid (or set of nucleic acids) encoding a full-length anti-TRAIL antibody is provided, including any of the full-length anti-TRAIL antibodies described herein. In some embodiments, a nucleic acid (or a set of nucleic acids) of an anti-TRAIL antibody described herein may also include a nucleic acid sequence encoding a polypeptide tag (e.g., a protein purification tag, his tag, HA tag).

Also contemplated herein are isolated host cells comprising an anti-TRAIL antibody, isolated nucleic acids encoding an anti-TRAIL antibody polypeptide component, or vectors comprising nucleic acids encoding an anti-TRAIL antibody polypeptide component described herein.

The application also includes variants of these nucleic acid sequences. For example, a variant comprises a nucleotide sequence that hybridizes under at least moderately stringent hybridization conditions to a nucleic acid sequence encoding an anti-TRAIL antibody of the application.

The application also provides vectors into which the nucleic acid sequences of the application can be inserted.

Briefly, an anti-TRAIL antibody (e.g., a full length anti-TRAIL antibody) may be expressed by inserting a natural or synthetic nucleic acid encoding the anti-TRAIL antibody into a suitable expression vector such that the nucleic acid is operably linked to 5' and 3' regulatory elements, including, for example, promoters (e.g., lymphocyte-specific promoters) and 3' untranslated regions (UTRs). The vectors may be suitable for replication and integration in eukaryotic host cells. Typical cloning and expression vectors contain transcriptional and translational terminators, initiation sequences, and promoters that regulate the expression of a nucleic acid sequence of interest.

The nucleic acids of the application can also be used for nucleic acid immunization and gene therapy by using standard gene delivery protocols. Nucleic acid delivery methods are known in the art. See, for example, U.S. Pat. nos.5,399,346, 5,580,859, 5,589,466, the entire contents of which are incorporated herein by reference. In some embodiments, the application also provides gene therapy vectors.

Nucleic acids can be cloned into many types of vectors. For example, the nucleic acid may be cloned into vectors including, but not limited to, plasmids, phagemids, phage derivatives, animal viruses and cosmids. Vectors of particular interest include expression vectors, replication vectors, probe-generating vectors and sequencing vectors.

In addition, the expression vector may be provided to the cell in the form of a viral vector. Viral vector technology is well known in the art and is described, for example, in Green and Sambrook(2013,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York), and other virology or molecular biology manuals. Viruses that may be used as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses. In general, suitable vectors include an origin of replication, promoter sequences, convenient restriction enzyme sites, and one or more selectable markers that function in at least one organism (see, e.g., WO 01/96584; WO 01/29058; and U.S. Pat.No.6,326,193).

Many virus-based systems have been developed for transferring genes into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. The selected gene may be inserted into a vector and packaged into retroviral particles using techniques known in the art. The recombinant virus is then isolated and delivered to cells of the subject in vivo or in vitro. Many retroviral systems are known in the art. In some embodiments, an adenovirus vector is used. Many adenoviral vectors are known in the art. In some embodiments, lentiviral vectors are used. Retroviral-derived vectors, such as lentiviruses, are suitable tools for achieving long-term gene transfer, as they allow for long-term stable integration of the transgene and propagation in daughter cells. Lentiviral vectors have additional advantages over retroviruses derived from tumors, such as the mouse leukemia virus, in that they can transduce non-dividing cells, such as hepatocytes. At the same time, it has the additional advantage of low immunogenicity.

Other promoter elements, e.g., enhancers, regulate the transcription initiation frequency. Typically they are located 30-110bp upstream of the start site, although many promoters have recently been found to contain functional elements downstream of the start site as well. The spacing between promoter elements is generally flexible so that the function of the promoter is maintained when the elements are interchanged or moved in position relative to each other. In the thymidine kinase (tk) promoter, the increase in the spacing between promoter elements to 50bp activity begins to decrease.

One example of a suitable promoter is the immediate early Cytomegalovirus (CMV) promoter sequence. The promoter sequence is a strong constitutive promoter sequence capable of driving high levels of expression of any polynucleotide sequence operably linked thereto. Another example of a suitable promoter is the elongation factor 1 alpha (EF-1 alpha) promoter. However, other constitutive promoters may also be used, including but not limited to simian virus 40 (SV 40) early promoter, mouse Mammary Tumor Virus (MMTV), human immunodeficiency virus long terminal repeat (HIV-LTR) promoter, moMuLV promoter, avian leukemia virus promoter, epstein-Barr virus immediate early promoter, rous sarcoma virus promoter, and human gene promoters including, for example, but not limited to, actin promoter, myosin promoter, hemoglobin promoter, and creatine kinase promoter. Furthermore, the application should not be limited to the use of constitutive promoters alone, and inducible promoters are also contemplated by the application. The use of an inducible promoter provides a molecular switch that enables expression of the polynucleotide sequence to which it is operably linked when such expression is desired and turns off expression when not desired. Inducible promoters include, but are not limited to, metallothionein promoters, glucocorticoid promoters, progesterone promoters, and tetracycline promoters.

In some embodiments, expression of the anti-TRAIL antibody is inducible. In some embodiments, the nucleic acid sequence encoding an anti-TRAIL antibody is operably linked to an inducible promoter, including any of the inducible promoters described herein.

Inducible promoter

The use of an inducible promoter provides a molecular switch that can initiate expression of a polynucleotide sequence operably linked thereto when expression is desired and which can shut down expression when expression is not desired. Exemplary inducible promoters suitable for use in eukaryotic cells include, but are not limited to, hormone regulatory elements (see, e.g., mader, S.and White, J.H. (1993) Proc.Natl. Acad.Sci.USA 90:5603-5607), synthetic ligand regulatory elements (see, spencer, D.M.et al (1993) Science 262:1019-1024), and ionizing radiation regulatory elements (see, Manome,Y.et al.(1993)Biochemistry 32:10607-10613;Datta,R.et al.(1992)Proc.Natl.Acad.Sci.USA 89:1014-10153). other exemplary inducible promoters suitable for use in mammalian systems in vivo or in vitro see GINGRICH ET al. (1998) Annual Rev. Neurosci 21:377-405. In some embodiments, the inducible promoter system for expression of anti-TRAIL antibodies is the Telact system, in some embodiments, the inducible promoter system for expression of anti-TRAIL antibodies is the E.coli lac inhibitory system.

One exemplary inducible promoter system employed in the present application is the Tet system. The system is based on the Tet system described by golden et al (1993). In one exemplary embodiment, the target polynucleotide is controlled by a promoter comprising one or more Tet operator (TetO) sites. In the inactive state, the Tet repressor (TetR) binds to the TetO site and inhibits transcription of the promoter. In the activated state, for example, in the presence of an inducer such as tetracycline (Tc), anhydrous tetracycline, doxycycline (Dox), or an active analog thereof, the inducer will release TetR from TetO, resulting in transcription. Doxycycline is a member of the tetracycline antibiotic family under the chemical name 1-dimethylamino-2, 4a,5, 7-pentahydroxy-11-methyl-4, 6-dioxo-1, 4a,11 a,12 a-hexahydrotetraene-3-carboxamide.

In one embodiment, tetR is codon optimized for expression in mammalian cells, such as mouse or human cells. Because of the degeneracy of the genetic code, most amino acids are encoded by more than one codon, such that the sequence of a given nucleic acid has a large number of variants without any change in the amino acid sequence encoded thereby. However, many organisms differ in codon usage, also known as "codon preference" (i.e., the preference of a given amino acid to use a particular codon). Codon preference is generally related to the presence of dominant tRNA species for a particular codon, which in turn increases the efficiency of mRNA translation. Coding sequences derived from a particular species (e.g., prokaryotes) can thus be tailored by codon optimization to enhance their expression in a different species (e.g., eukaryotes).

Other specific variations of the Tet system include the following "Tet-Off" and "Tet-On" systems. In the Tet-off system, transcription is inactive in the presence of Tc or Dox. In this system, the tetracycline-regulated transcriptional activator protein (tTA), consisting of TetR fused to the strong transcriptional activation domain of the herpes simplex virus VP16, regulates expression of the target nucleic acid under the transcriptional control of the tetracycline responsive promoter element (TRE). The TRE element consists of a TetO sequence tandem fused to a promoter (typically the smallest promoter sequence derived from the human cytomegalovirus immediate early promoter). In the absence of Tc or Dox, tTA binds to TRE and activates transcription of the target gene. In the presence of Tc or Dox, tTA cannot bind to TRE and the target gene cannot be expressed.

In contrast, in the Tet-On system, transcription is active in the presence of Tc or Dox. The Tet-On system is based On the reverse tetracycline regulated transcriptional activator rtTA. Like tTA, rtTA is a fusion protein consisting of the TetR repressor and VP16 transcriptional activation domain. However, a 4 amino acid change in the DNA binding region of TetR alters the binding properties of rtTA such that it recognizes only the tetO sequence on the target transgenic TRE in the presence of Dox. Therefore in the Tet-On system rtTA activates the transcription of the target gene regulated by TRE only in the presence of Dox.

Another inducible promoter system is the E.coli lac repressor system (see Brown et al, cell 49:603-612 (1987)). The Lac repressor system functions by regulating transcription of a polynucleotide of interest operably linked to a promoter comprising the Lac operator (lacO). The Lac repressor (lacR) binds to LacO and thereby prevents transcription of the target polynucleotide. Expression of the polynucleotide of interest is induced by a suitable inducer, for example isopropyl- β -D thiogalactopyranoside (IPTG).

To assess the expression of the polypeptide or portion thereof, the expression vector to be introduced into the cell may further comprise a selectable marker gene or a reporter gene or both to facilitate identification and selection of the expressing cell from a population of cells transfected or infected with the viral vector. In other aspects, the selectable marker may be carried on separate DNA fragments and used in a co-transfection experiment. Either the selectable marker gene or the reporter gene may be flanked by appropriate regulatory sequences to enable expression in the host cell. Useful selectable markers include, for example, antibiotic resistance genes, such as neo and the like.

Reporter genes can be used to identify potentially transfected cells and evaluate the function of regulatory sequences. Typically, a reporter gene is a gene that is not present in or expressed by a recipient organism or tissue, and encodes a polypeptide whose expression is manifested by some readily detectable property, such as enzymatic activity. After the DNA is introduced into the recipient cell, the expression of the reporter gene is detected at an appropriate time. Suitable reporter genes may include genes encoding luciferases, beta-galactosidases, chloramphenicol acetyl transferase, secreted alkaline phosphatase, or green fluorescent protein (see Ui-Tel et al, 2000FEBS Letters 479:79-82). Suitable expression systems are well known and may be prepared by known techniques or obtained commercially. In general, constructs that display the smallest 5' flanking region of the highest expression level of the reporter gene are considered promoters. Such promoter regions may be linked to reporter genes and used to assess the ability of certain substances to regulate promoter-driven transcription.

In some embodiments, nucleic acids encoding any of the full-length anti-TRAIL antibodies described herein are provided. In some embodiments, the nucleic acid comprises one or more nucleic acid sequences encoding a full length anti-TRAIL antibody heavy and light chain. In some embodiments, each of the one or more nucleic acid sequences is contained in a separate vector. In some embodiments, at least some of the nucleic acid sequences are contained in the same vector. In some embodiments, all nucleic acid sequences are contained in the same vector. The vector may be selected from, for example, mammalian expression vectors and viral vectors (such as vectors derived from retroviruses, adenoviruses, adeno-associated viruses, herpesviruses and lentiviruses).

Methods for introducing and expressing genes into cells are known in the art. In the context of expression vectors, the vector may be readily introduced into a host cell, such as a mammalian cell, bacterial, yeast or insect cell, by any method known in the art. For example, the expression vector may be introduced into the host cell by physical, chemical or biological means.

Physical methods for introducing polynucleotides into host cells include calcium phosphate precipitation, lipofection, gene gun methods, microinjection, electroporation, and the like. Methods for preparing cells comprising vectors and/or exogenous nucleic acids are well known in the art. See, e.g., Green and Sambrook(2013,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York). in some embodiments, the polynucleotide is introduced into the host cell by calcium phosphate transfection.

Biological methods for introducing polynucleotides of interest into host cells include the use of DNA and RNA vectors. Viral vectors, particularly retroviral vectors, have become the most widely used method for inserting genes into mammalian cells, such as human cells. Other viral vectors may be derived from lentiviruses, poxviruses, herpes simplex virus type 1, adenoviruses, adeno-associated viruses, and the like. See, e.g., U.S. Pat. nos.5,350,674 and 5,585,362.

Chemical methods for introducing polynucleotides into host cells include colloidal dispersion systems, such as macromolecular complexes, nanocapsules, microspheres, magnetic beads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system used as a delivery vehicle in vivo and in vitro is a liposome (e.g., an artificial membrane vesicle).

In the case of non-viral delivery systems, an exemplary delivery vehicle is a liposome. The use of lipid formulations to introduce nucleic acids into host cells (in vitro, ex vivo or in vivo) is contemplated. In another aspect, the nucleic acid may be conjugated to a lipid. The lipid-bound nucleic acid may be entrapped within the aqueous interior of the liposome, dispersed within the lipid bilayer of the liposome, linked to the liposome by a linking molecule that binds to the liposome and the oligonucleotide, entrapped in the liposome, formed a complex with the liposome, dispersed in a solution containing the lipid, mixed with the lipid, bound to the lipid, suspended in the lipid, contained in or mixed with the micelle, or otherwise bound to the lipid. The lipid, lipid/DNA or lipid/expression vector-related composition is not limited to any particular structure in solution. For example, they may exist in a bilayer structure, in micelles, or in a "collapsed" structure. They may also be simply dispersed in solution, possibly forming aggregates of non-uniform size or shape. Lipids are fatty substances, which may be naturally occurring or synthetic. For example, lipids include fat droplets naturally occurring in the cytoplasm, as well as a class of compounds containing long chain aliphatic hydrocarbons and derivatives thereof, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes.

Regardless of the method used to introduce exogenous nucleic acid into a host cell or otherwise expose the cell to the inhibitors of the application, various experiments can be performed in order to confirm the presence of the recombinant DNA sequence in the host cell. Such assays include, for example, "molecular biology" assays well known to those of skill in the art. For example Southern and Northern blotting, RT-PCR and PCR; "biochemical" assays, such as detecting the presence or absence of a particular polypeptide, such as by immunological methods (ELISAs and Western blots) or by the assays described herein, fall within the scope of the application.

Preparation of anti-TRAIL antibodies

In some embodiments, the anti-TRAIL antibody is a monoclonal antibody or is derived from a monoclonal antibody. In some embodiments, the anti-TRAIL antibody comprises V _H and V _L, or variants thereof, from a monoclonal antibody. In some embodiments, the anti-TRAIL antibody further comprises CH1 and CL regions from a monoclonal antibody, or a variant thereof. Monoclonal antibodies can be prepared using methods known in the art, including hybridoma cell methods, phage display methods, or using recombinant DNA methods, for example. Furthermore, exemplary phage display methods are described herein and in the examples below.

In hybridoma cell methods, hamsters, mice, or other suitable host animals are typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Or lymphocytes may be immunized in vitro. The immunizing agent may include a polypeptide or fusion protein of the protein of interest. Typically, peripheral Blood Lymphocytes (PBLs) are used if human cells are desired, whereas spleen cells or lymph node cells are used if non-human mammalian cells are desired. Lymphocytes are fused with an immortalized cell line, such as polyethylene glycol, using an appropriate fusion agent to form a hybridoma cell. Immortalized cell lines are typically transformed mammalian cells, especially myeloma cells of rodent, bovine and human origin. Rat or mouse myeloma cell lines are typically employed. The hybridoma cells may be cultured in a suitable medium, which preferably contains one or more substances that inhibit the growth or survival of the unfused immortalized cells. For example, if the parent cell lacks hypoxanthine-guanine phosphoribosyl transferase (HGPRT or HPRT), the hybridoma cell culture medium typically includes hypoxanthine, aminopterin, and thymidine (HAT medium), which prevents HGPRT-deficient cells from growing.

In some embodiments, the immortalized cell lines fuse efficiently, ensure high levels of stable expression of antibodies by the antibody-producing cell of choice, and are sensitive to certain media, such as HAT media. In some embodiments, the immortal cell line is a mouse myeloma cell line, available from, for example, the sork cell collection in san diego, california and the american type culture collection in ma, virginia. Human myeloma and murine-human hybrid myeloma cell lines are also described for use in the production of humanized monoclonal antibodies.

The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the polypeptide. The binding specificity of monoclonal antibodies produced by hybridoma cells can be determined by immunoprecipitation or in vitro binding assays, such as Radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques or analytical methods are known in the art. The binding affinity of a monoclonal antibody can be determined by, for example, the Scatchard (Scatchard) assay described in Munson and Pollard, anal. Biochem.,107:220 (1980).

After the desired hybridoma cells are identified, the target clone may be subcloned by limiting dilution and cultured by standard methods. Suitable media for this purpose include, for example, modified Eagle Medium (DMEM) and RPMI-1640 medium. Alternatively, the hybridoma cells may be grown as ascites in a mammal.

Monoclonal antibodies secreted by subclones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification methods, such as protein A-sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

In some embodiments, according to any of the anti-TRAIL antibodies described herein, the anti-TRAIL antibody comprises a sequence selected from the group consisting of clones of an antibody library (e.g., a phage library displaying scFv or Fab fragments). The clones may be identified by screening combinatorial libraries of antibody fragments having the desired activity. For example, a variety of methods are known in the art for generating phage display libraries and screening these libraries to obtain antibodies of the desired binding characteristics. These methods are reviewed in Hoogenboom et al.,Methods in Molecular Biology 178:1-37(O'Brien et al.,ed.,Human Press,Totowa,N.J.,2001), for example, and further described in McCafferty et al.,Nature 348:552-554;Clackson et al.,Nature 352:624-628(1991);Marks et al.,J.Mol.Biol.222:581-597(1992);Marks and Bradbury,Methods in Molecular Biology 248:161-175(Lo,ed.,Human Press,Totowa,N.J.,2003);Sidhu et al.,J.Mol.Biol.338(2):299-310(2004);Lee et al.,J.Mol.Biol.340(5):1073-1093(2004);Fellouse,Proc.Natl.Acad.Sci.USA 101(34):12467-12472(2004);and Lee et al.,J.Immunol.Methods 284(1-2):119-132(2004), for example.

In some phage display methods, all components of the V _H and V _L genes are cloned separately by Polymerase Chain Reaction (PCR) and randomly recombined in a phage library, followed by screening for phage capable of binding to antigen, as described in Winter et al, ann.Rev.Immunol.,12:433-455 (1994). Phage typically display antibody fragments as scFv fragments or as Fab fragments. The immune-derived library phage provides high affinity antibodies to the immunogen without the need to construct hybridoma cells. Alternatively, a natural repertoire (e.g., from a human) can be cloned to provide a single antibody source against multiple non-self and self-antigens without any immunization, as described in GRIFFITHS ET al, EMBO J,12:725-734 (1993). Finally, natural libraries can also be prepared by cloning non-rearranged V-gene fragments from stem cells and encoding CDR3 hypervariable regions using PCR primers comprising random sequences and completing the rearrangement in vitro, as described in Hoogenboom AND WINTER, J.mol.biol.,227:381-388 (1992). Patent publications describing human antibody phage libraries include, for example, U.S. Pat. nos. 5,750,373 and US Patent Publication Nos.2005/0079574、2005/0119455、2005/0266000、2007/0117126、2007/0160598、2007/0237764、2007/0292936 and 2009/0002360.

The anti-TRAIL antibodies are prepared by a method of phage display screening of the portion of the library of anti-TRAIL antibodies that specifically bind to the target TRAIL. The library may be a human scFv phage display library, having at least 1 x 10 ⁹ (e.g., at least 1×10⁹、2.5×10⁹、5×10⁹、7.5×10⁹、1×10¹⁰、2.5×10¹⁰、5×10¹⁰、7.5×10¹⁰ or 1 x 10 ¹¹) diversity of unique human antibody fragments. In some embodiments, the library is a human natural library constructed from DNA extracted from PMBCs and spleen of healthy subjects, comprising all human heavy and light chain subfamilies. In some embodiments, the library is a human natural library constructed from DNA extracted from PMBCs isolated from patients with various diseases, such as patients with autoimmune diseases, cancer patients, and patients with infectious diseases. In some embodiments, the library is a semi-synthetic human library in which the heavy chain CDR3 is entirely random, with all amino acids (except cysteine) present at any given position with the same probability. (see, e.g., hoet, R.M. et al, nat. Biotechnol.23 (3): 344-348, 2005). In some embodiments, the heavy chain CDR3 of the semi-synthetic human library is between 5 and 24 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24) amino acids in length. In some embodiments, the library is a fully synthetic phage display library. In some embodiments, the library is a non-human phage display library.

Phage clones with high affinity for target TRAIL may be screened by iterative binding of phage to target TRAIL bound to a solid support (e.g. beads for solution panning or mammalian cells for cell panning), followed by removal of unbound phage and elution of specifically bound phage. The bound phage clones are then eluted and used to infect appropriate host cells, e.g., E.coli XL1-Blue, for expression and purification. Phage clones that specifically bind TRAIL can be enriched by multiple rounds of panning (e.g., 2,3, 4, 5, 6 or more rounds), such as solution panning, cell panning, or both. Specific binding of the enriched phage clones to the target TRAIL can be detected by any method known in the art, including, for example, ELISA and FACS.

Another method of screening antibody libraries is to display proteins on the surface of yeast cells. Wittrup et al (U.S. Pat. Nos. 6,699,658 and 6,696,251) developed a method for displaying libraries of yeast cells. In this yeast display system, one component comprises a yeast lectin protein (Aga 1) anchored to the yeast cell wall, and the other component comprises a second subunit of lectin protein Aga2, which subunit can bind to the Aga1 protein via disulfide bonds and thus be displayed on the yeast cell surface. The Aga1 protein is expressed by integrating the Aga1 gene into the yeast chromosome. A library of single-chain variable fragments (scFv) was fused to the Aga2 gene in a yeast display plasmid, and after transformation, the library was retained in the yeast due to the presence of additional nutritional markers. Both the Aga1 and Aga2 proteins are expressed under the control of a galactose-inducible promoter.

The human antibody V gene library (fragments V _H and V _K) was obtained by PCR using a degenerate set of primers (Sblattero, D.and Bradbury, A.immunotechnology 3,271-278 1998). PCR templates were derived from commercially available RNA or cDNA, including PBMC, spleen, lymph nodes, bone marrow and tonsils. Independent V _H and V _K PCR libraries were pooled and assembled into scFv forms by overlap extension PCR (Sheets, M.D.et al, proc.Natl. Acad.Sci. USA 95,6157-6162 1998). To construct a yeast scFv display library, the resulting scFv PCR product was cloned into a yeast display plasmid in yeast by homologous recombination.

(Chao,G,et al,Nat Protoc.2006;1(2):755-68.Miller KD,et al.Current Protocols in Cytometry 4.7.1-4.7.30,2008).

Anti-TRAIL antibodies may be screened using a mammalian cell display system in which the antibody portion is displayed on the cell surface and antibodies specifically targeting TRAIL are isolated by antigen-directed screening methods (as described in U.S. patent No.7,732,195B2). A Chinese Hamster Ovary (CHO) cell library displaying a large number of human IgG antibody genes can be established and used to discover clones expressing high affinity antibody genes. Another display system has been developed that allows the same protein to be displayed and secreted simultaneously on the cell surface by alternative splicing, wherein the displayed protein phenotype remains genotype-dependent, allowing the secreted soluble antibody to be characterized simultaneously in biophysical and cell function-based assays. This method overcomes many of the limitations previously exhibited by mammalian cells and enables direct screening and maturation of antibodies in the form of full-length, glycosylated IgGs (Peter M.Bowers, et al Methods 2014, 65:44-56). Transient expression systems are suitable for single round antigen selection prior to antibody gene recovery and are therefore most useful for selecting antibodies from smaller libraries. Stable exon vectors offer an attractive option. The exon vectors can be transfected efficiently and stably maintained at low copy numbers, allowing multiple rounds of panning and resolution of more complex antibody libraries.

The IgG library was constructed based on ligation of germline sequence V gene segments isolated from a population of human donors with rearranged (D) J regions. RNA collected from 2000 human blood samples was reverse transcribed into cDNA, and V _H and V _K fragments were amplified using V _H and V _K specific primers and purified by gel extraction. The V _H and V _K fragments were subcloned into a display vector comprising an IgG1 or K constant region, respectively, and then electroporated or transduced 293T into cells to prepare an IgG library. To prepare the scFv antibody display library, V _H and V _K were linked to generate scFv, which were then subcloned into display vectors and electroporated or transduced 293T cells. It is well known that IgG libraries are constructed based on germline sequence V gene segments and rearranged (D) J regions isolated from a population of donors, which may be mice, rats, rabbits or monkeys.

Monoclonal antibodies can also be prepared by recombinant DNA methods, for example as described in U.S. patent No.4,816,567. The DNA encoding the monoclonal antibodies of the application can be readily isolated and sequenced by conventional methods, such as by oligonucleotide probes that specifically bind to the light and heavy chain genes encoding murine antibodies. Hybridoma cells as described above or TRAIL-specific phage clones of the application may be used as a source of such DNA. After isolation, the DNA may be placed in an expression vector, which is then transfected into a host cell, such as simian COS cells, chinese hamster ovary Cancer (CHO) cells, or myeloma cells that do not produce immunoglobulins, to obtain monoclonal antibodies synthesized in the recombinant host cell. The DNA may also be modified, for example by replacing the human heavy and light chain constant regions with coding sequences and/or by replacing homologous non-human sequences with framework regions (U.S. patent No.4,816,567; morrison et al, supra), or by covalently joining all or part of the coding sequence of an immunoglobulin to the coding sequence of a non-immunoglobulin polypeptide. Such non-immunoglobulin polypeptides may replace the constant regions of the antibodies of the application, or may replace an antigen binding site in the variable domains of the antibodies of the application, to form chimeric bivalent antibodies.

The antibody may be a monovalent antibody. Methods of making monovalent antibodies are known in the art. For example, a recombinant expression method involving an immunoglobulin light chain and a modified heavy chain. Heavy chains are typically truncated at any position in the Fc region to prevent heavy chains from cross-linking with each other. Or the relevant cysteine residues are substituted with other amino acid residues or deleted to prevent cross-linking.

In vitro methods are also suitable for the preparation of monovalent antibodies. Digestion of antibodies to produce antibody fragments, particularly Fab fragments, may be accomplished using any method known in the art.

The antibody variable domain having the desired binding specificity (antibody-antigen binding site) may be fused to an immunoglobulin constant region. Preferably fusion with an immunoglobulin heavy chain constant region, which comprises at least part of the hinge, CH2 and CH3 regions. In some embodiments, the first heavy chain constant region (CH 1) comprising the necessary site for light chain binding is present in at least one fusion. The DNA encoding the immunoglobulin heavy chain fusion, and if desired, the immunoglobulin light chain, is inserted into a separate expression vector and co-transfected into a suitable host organism.

Fully human and humanized antibodies

The anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) may be a fully human antibody or a humanized antibody. Humanized forms of non-human (e.g., mouse) antibody portions are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (e.g., fv, fab, fab ', F (ab') ₂, scFv or other antigen-binding subsequences of antibodies) that typically include minimal sequences derived from non-human immunoglobulins. Humanized antibodies include human immunoglobulins, immunoglobulin chains or fragments thereof (recipient antibodies) in which residues from a recipient CDR are replaced by non-human (donor antibody) CDR residues having the desired specificity, affinity and properties, such as mouse, rat or rabbit CDRs. In some embodiments, the human immunoglobulin Fv framework region residues are replaced by corresponding non-human residues. Humanized antibodies may also comprise amino acid residues that are neither of the recipient antibody nor in the introduced CDR or framework sequences. Typically, a humanized antibody comprises at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are human immunoglobulin consensus sequences.

Typically, humanized antibodies contain one or more amino acid residues introduced from a non-human source. Those non-human amino acid residues are often referred to as "import" residues, typically from "import" variable domains. According to some embodiments, humanization may be performed substantially as described below by Winter and colleagues (Jones et al.,Nature,321:522-525(1986);Riechmann et al.,Nature,332:323-327(1988);Verhoeyen et al.,Science,239:1534-1536(1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Thus, this "humanized" antibody portion (U.S. patent No.4,816,567), which is substantially less than a fully human antibody, has its variable domains replaced by corresponding sequences from a non-human source. In practice, humanized antibody portions are typical human antibody portions in which some CDR residues and possibly some framework region residues are replaced with residues from similar sites in rodent antibodies.

Fully human antibodies are an alternative to humanization. For example, transgenic animals (e.g., mice) that are capable of producing a complete fully human antibody library after immunization without endogenous immunoglobulin production can now be prepared. For example, homozygous deletion of the antibody heavy chain Junction (JH) gene in chimeric and germ-line mutant mice has been reported to completely suppress endogenous antibody production. Transfer of an array of human germline immunoglobulin genes into such germline mutant mice can result in the production of human antibodies under antigenic stimulation, see, e.g., akobovits et al.,PNAS USA,90:2551(1993);Jakobovits et al.,Nature,362:255-258(1993);Bruggemann et al.,Year in Immunol.,7:33(1993);U.S.Patent Nos.5,545,806,5,569,825,5,591,669,5,545,807; and WO 97/17852. Alternatively, fully human antibodies can be prepared by introducing a human immunoglobulin locus into a transgenic animal (e.g., a mouse in which endogenous immunoglobulin genes have been partially or fully silenced). Upon antigen stimulation, the production of fully human antibodies can be found to be very similar in all respects to their production in humans, including gene rearrangement, assembly and antibody libraries. Such a method is described, for example, in U.S.Patent Nos.5,545,807;5,545,806;5,569,825;5,625,126;5,633,425;and 5,661,016,and Marks et al.,Bio/Technology,10:779-783(1992);Lonberg et al.,Nature,368:856-859(1994);Morrison,Nature,368:812-813(1994);Fishwild et al.,Nature Biotechnology,14:845-851(1996);Neuberger,Nature Biotechnology,14:826(1996);Lonberg and Huszar,Intern.Rev.Immunol.,13:65-93(1995).

Fully human antibodies are also produced by in vitro activation of B cells (see U.S. patents 5,567,610and 5,229,275) or by using various techniques known in the art, including phage display libraries. Hoogenboom AND WINTER, J.mol.biol.,227:381 (1991); the techniques of Marks et al, J.mol.biol.,222:581 (1991), cole et al, and Boerner et al can also be used to prepare fully human monoclonal antibodies. See Cole et al.,Monoclonal Antibodies and Cancer Therapy,Alan R.Liss,p.77(1985)and Boerner et al.,J.Immunol.,147(1):86-95(1991).

Anti-TRAIL antibody variants

In some embodiments, the amino acid sequences of anti-TRAIL antibody variants provided herein (e.g., full length anti-TRAIL antibodies) are also contemplated. For example, it may be desirable to improve the binding affinity and/or other biological activity of antibodies. The amino acid sequence of an antibody variant may be prepared by introducing appropriate modifications in the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and/or insertions and/or substitutions of residues in the amino acid sequence of the antibody. The final construction can be accomplished by any combination of amino acid residue deletions, insertions, and substitutions to impart the desired characteristics. For example, antigen binding.

In some embodiments, anti-TRAIL antibody variants having one or more amino acid substitutions are provided. Target sites for substitution mutations include hypervariable regions (HVRs) and Framework Regions (FRs). Amino acid substitutions may be introduced into the antibody of interest to screen for products of a desired activity, e.g., improved biological activity, retention/improvement of antigen binding capacity, reduced immunogenicity, or improved ADCC or CDC.

Conservative substitutions are shown in table 4 below.

TABLE 4 conservative substitutions

Amino acids are classified into different classes according to the nature of the side chain:

a. Hydrophobic amino acid: norleucine Norleucine, methionine Met, alanine Ala, valine Val, leucine Leu, isoleucine Ile;

b. Neutral hydrophilic amino acid: cysteine Cys, serine Ser, threonine Thr, asparagine Asn, glutamine Gln;

c. Acidic amino acid: aspartic acid Asp, glutamic acid Glu;

d. Basic amino acid: histidine His, lysine Lys, arginine Arg;

e. Contains amino acids affecting the chain direction: glycine Gly, proline Pro;

f. aromatic amino acid: tryptophan Trp, tyrosine Tyr, phenylalanine Phe.

Substitutions of non-conservative amino acids include substitution of one of the above classes into another class.

One exemplary substitution variant is an affinity matured antibody, conveniently produced using, for example, phage display-based affinity maturation techniques. Briefly, one or more CDR residues are mutated, the variant antibody portion is displayed on a phage, and variants are screened for a particular biological activity (e.g., activity to inhibit apoptosis or increased antibody affinity). Alterations (e.g., substitutions) may be made in the HVRs regions to obtain improved antibody affinity or biological activity. The binding affinities of the resulting variants V _H and V _L can be tested for changes in the "hot spot" of the HVR, i.e. codon-encoded residues that undergo high frequency mutations during somatic maturation (see, e.g., chowdhury, methods mol. Biol.207:179-196 (2008)), and/or at Specific Determinant Residues (SDRs). Methods for constructing and reselecting affinity maturation from secondary libraries have been described in some literature, for example ,Hoogenboomet al.in Methods in Molecular Biology 178:1-37(O'Brien et al.,ed.,Human Press,Totowa,NJ,(2001)).

In some affinity maturation embodiments, diversity is introduced into the selected variable genes for affinity maturation by any of a variety of methods (e.g., error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then created. The library is screened to identify antibody variants with the desired affinity. Another approach to introducing diversity involves HVR-mediated approaches in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding are specifically recognized, for example, using alanine scanning mutagenesis or modeling. CDR-H3 and CDR-L3 regions are generally particularly important targets.

In some embodiments, substitutions, insertions, or deletions may occur within one or more HVRs, provided that such changes do not substantially reduce the ability of the antibody to bind to an antigen. For example, conservative changes (e.g., conservative substitutions provided herein) may be made in HVRs that do not substantially reduce binding affinity. These changes may occur outside the HVR "hot spot" or SDRs region. In some embodiments the variant V _H and V _L sequences provided above, each HVR is either unchanged or comprises no more than 1,2, or 3 amino acid substitutions.

One useful method by which amino acid residues or regions of an antibody that can be targeted for mutation can be identified is known as "alanine scanning mutagenesis" as described in Cunninghamand Wells (1989) Science, 244:1081-1085. In this method, one or a group of target residues (e.g., charged residues such as arginine, aspartic acid, histidine, lysine, and glutamic acid) are substituted with neutral or negatively charged amino acids (e.g., alanine or glutamic acid) to determine whether the interaction of the antibody with the antigen is affected. Substitutions may be further introduced at the amino acid position to demonstrate functional sensitivity of the position to the initial substitution. Alternatively or additionally, the contact site between the antibody and the antigen is identified by the crystal structure of the antigen-antibody complex. These contact site residues and adjacent residues may be targeted or eliminated as substitution candidates. Variants are screened to determine if they have the desired properties.

Insertion of amino acid sequences, including fusion at the amino and/or carboxy terminus, ranges in length from 1 residue to polypeptides comprising 100 or more residues, and also includes insertion of 1 or more amino acid residues within the sequence. Examples of terminal insertions include antibodies having a methionyl residue at the N-terminus. Other insertional variants of antibody molecules include polypeptides that fuse an enzyme (e.g., ADEPT) or increase the serum half-life of an antibody at the N-or C-terminus of the antibody molecule.

Variant Fc region

In some embodiments, one or more amino acid modifications are introduced into the Fc region of an antibody described herein (e.g., a full length anti-TRAIL antibody or an anti-TRAIL antibody fusion protein), thereby producing an Fc region variant. In some embodiments, the Fc region variant has enhanced ADCC potency, typically associated with Fc-binding receptors (FcRs). In some embodiments, the Fc region variant has reduced ADCC potency. There are many examples of alterations or mutations in Fc sequences affecting their potency, for example, WO 00/42072 and SHIELDS ET al J biol. Chem.9 (2): 6591-6604 (2001) describe antibody variants with increased or decreased binding to FcRs. The contents of these publications are incorporated herein by reference.

Antibody-dependent cell-mediated cytotoxicity (ADCC) is the mechanism of action of therapeutic antibodies against tumor cells. ADCC is a cell-mediated immune defense in which effector cells of the immune system actively lyse target cells (e.g., cancer cells) when antigens on the surface of the target cell membrane are bound by specific antibodies (e.g., anti-TRAIL antibodies). Typically ADCC effects involve NK cells activated by antibodies. NK cells express the Fc receptor CD16. The receptor recognizes and binds to the Fc portion of an antibody molecule that binds to the surface of a target cell. The most common Fc receptor on the surface of NK cells is CD16 or fcyriii. Binding of the Fc receptor to the Fc region of the antibody results in activation of NK cells, releasing the cell lysis particles, followed by apoptosis of the target cells. The killing of tumor cells by ADCC can be determined by experiments specific for NK-92 cells transfected with high affinity FcR. The results were compared with wild-type NK-92 which did not express FcR.

In some embodiments, the application also provides anti-TRAIL antibody variants (e.g., full length anti-TRAIL antibody variants) comprising an Fc region having a portion, but not all, of effector function such that it has an extended half-life in vivo, whereas specific effector functions (e.g., CDC or ADCC) are not necessary or detrimental, which anti-TRAIL antibodies are desirable candidates for the application. Reduction/elimination of CDC and/or ADCC activity is confirmed by cytotoxicity assays in vitro and/or in vivo. For example, antibodies were confirmed to lack fcγr binding capacity (and thus potentially ADCC activity) by an Fc receptor (FcR) binding assay but still retain FcRn binding capacity. Among the major cells mediating ADCC, NK cells express fcyriii only, whereas monocytes express fcyri, fcyrii and fcyriii. The expression of FcR on hematopoietic cells is summarized in Table 3 at page 464 of RAVETCHAND KINET Annu. Rev. Immunol.9:457-492 (1991). Non-limiting examples of in vitro evaluation of ADCC activity of a target molecule are described in U.S. Pat. No.5,500,362 (see, e.g., Hellstrom,I.et al.Proc.Nat'l Acad.Sci.USA 83:7059-7063(1986)and Hellstrom,I et al.,Proc.Nat'l Acad.Sci.USA 82:1499-1502(1985);U.S.Pat.No.5,821,337(see Bruggemann,M.et al.,J.Exp.Med.166:1351-1361(1987)). or non-radioactive cytotoxicity assays (see, e.g., ACTI ^TM flow cytometry, calif.) and CYTOTOX 96 ^TM non-radioactive cytotoxicity assays (Promega, madison, wis.)) such assays employ effector cells including Peripheral Blood Mononuclear Cells (PBMC) and natural killer cells (NK.). Or alternatively, ADCC activity of a target molecule is detected in vivo, e.g., in animal models such as those described in Clynes et al, proc. Nat 'l acad. Sci. Usa 95:652-656 (1998.) while C1q binding assays can also be performed to confirm that antibodies cannot bind to C1q, thereby lacking CDC activity, see, e.g., WO 029879 and WO 2006/100402, C1q and C3C activation in order to evaluate complement activation (see, e.g., sank. 35:163:202) in animal models; cragg, M.S. et al, blood 101:1045-1052 (2003), and Cragg, M.S. and M.J. Glennie, blood103:2738-2743 (2004)), fcRn binding and in vivo clearance/half-life are determined using methods known in the art (see, e.g., petkova, S.B.et al, int' l.Immunol.18 (12): 1759-1769 (2006)).

Antibodies with reduced effector function comprising substitution of one or more residues at residues 238, 265, 269, 270, 297, 327 and 329 of the Fc region (u.s.pat.no. 6,737,056). These Fc variants include Fc variants with substitution of two or more residues at positions 265, 269, 270, 297 and 327, including Fc variants known as "DANA" with substitution of alanine at residues 265 and 297 (u.s.pat. No.7,332, 581).

Such antibody variants with increased or decreased binding to FcRs have been described (see, e.g., U.S. Pat.No.6,737,056; WO 2004/056312, and SHIELDS ET al, J.biol.chem.9 (2): 6591-6604 (2001)).

In some embodiments, an anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) variant is provided that comprises an Fc region variant having one or more amino acid substitutions capable of enhancing ADCC effect. In some embodiments, the Fc region variant comprises one or more amino substitutions at positions 298, 333, and/or 334 (EU residue numbering) of the Fc region that are capable of enhancing ADCC effects. In some embodiments, the anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) variant comprises amino acid substitutions at positions S298A, E333A, and K334A of the Fc region.

In some embodiments, the change in the Fc region results in a change (i.e., an increase or decrease) in C1q binding and/or Complement Dependent Cytotoxicity (CDC), as described in U.S.Pat.No.6,194,551, WO/51642, and Idusogie et al, J.Immunol.164:4178-4184 (2000).

In some embodiments, an anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) variant is provided that comprises an Fc region variant having one or more amino acid substitutions that is capable of extending half-life or enhancing binding to an Fc receptor (FcRn). Antibodies with extended half-life and improved FcRn binding are described in US2005/0014934A1 (hiton et al). These antibody Fc regions comprise one or more amino acid substitutions that enhance the binding of the Fc region to FcRn. These Fc variants comprise one or more substitutions in residues 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434 in the Fc region, for example a substitution in residue 434 in the Fc region (u.s.pat. No.7,371,826).

See also Duncan & Winter, nature 322:738-40 (1988); U.S. Pat. nos. 5,648,260; examples of other Fc region variants are provided in U.S. Pat. No.5,624,821 and WO 94/29351.

The application contemplates anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) comprising any one or a combination of the Fc variants described herein.

Glycosylation variants

In some embodiments, an anti-TRAIL antibody provided herein (e.g., a full length anti-TRAIL antibody) is altered to increase or decrease the degree of glycosylation of the anti-NGF antibody. The addition or deletion of glycosylation sites on anti-TRAIL antibodies may be conveniently accomplished by altering the amino acid sequence of the anti-NGF antibody or polypeptide portion thereof to thereby add or remove one or more glycosylation sites.

Wherein the anti-TRAIL antibody comprises an Fc region to which a saccharide may be linked. Natural antibodies produced by mammalian cells typically comprise branched double-antennary oligosaccharides, which are typically linked to the Fc region CH2 domain Asn297 via an N-linkage, see, e.g., wright et al, TIBTECH 15:26-32 (1997). The oligosaccharides may comprise a variety of sugars, such as mannose, N-acetylglucosaminide (GlcNAc), galactose and sialic acid, as well as trehalose attached to the GlcNAc of the "stem" of the double-antennary oligosaccharide structure. In some embodiments, the anti-TRAIL antibodies of the application may be oligosaccharide modified to produce anti-TRAIL antibody variants with certain improved properties.

N-glycans attached to the CH2 domain of the Fc region are heterogeneous. Antibodies or Fc fusion proteins produced in CHO cells are fucosylated by fucosyltransferase activity, see Shoji-Hosaka et al, J.biochem.2006,140:777-83. Typically, a small fraction of naturally occurring nonfucosylated IgGs can be detected in human serum. N-glycosylation of the Fc region is important for its binding to fcγr; whereas non-fucosylated N-glycans enhance the binding capacity of Fc to fcγriiia. Enhanced binding to FcRIIIa results in enhanced ADCC effect, which is advantageous in certain antibody therapeutic applications requiring cytotoxicity.

In some embodiments, enhanced effector function may be detrimental when Fc-mediated cellular cytotoxicity is not required. In some embodiments, the Fc fragment or CH2 domain is non-glycosylated. In some embodiments, glycosylation is prevented by mutating the N-glycosylation site in the CH2 domain.

In some embodiments, anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) variants are provided that comprise an Fc region, wherein the saccharide structure linked to the Fc region has reduced fucose or lacks fucose, which may enhance ADCC function. In particular, provided herein are anti-TRAIL antibodies having reduced fucose relative to the same anti-TRAIL antibodies produced by wild-type CHO cells. That is, they are characterized by having a smaller amount of fucose than antibodies produced by natural CHO cells (e.g., CHO cells producing a naturally glycosylated form, CHO cells containing the natural FUT8 gene). In some embodiments, the N-linked glycans of the anti-TRAIL antibody have less than 50%, 40%, 30%, 20%, 10% or 5% fucose. For example, the anti-TRAIL antibody may have a fucose content of 1% -80%, 1% -65%, 5% -65% or 20% -40%. In some embodiments, the N-linked glycans of the anti-TRAIL antibody do not comprise fucose, i.e., wherein the anti-TRAIL antibody is completely free of fucose, or is free of fucose or is defucosylated. The fucose content is determined by calculating the average fucose content in the sugar chains attached to Asn297 relative to the total amount of all sugar structures attached to Asn297 (e.g. complex, hybrid or mannose structures) as measured by MALDI-TOF mass spectrometry, as described in WO 2008/077546. Asn297 refers to the asparagine residue at position 297 of the Fc region (EU Fc region residue numbering system). However, asn297 may also be located upstream or downstream of position 297 by ±3 amino acids, i.e. between positions 294 and 300, due to minor sequence variations of the antibody. These fucosylated variants may have enhanced ADCC function. See, for example, US Patent publications US2003/0157108 (Presta, l.), US2004/0093621 (Kyowa Hakko Kogyo co., ltd). Examples of publications related to antibody variants that are "defucosylated" or "fucose deficient" include ,US2003/0157108;WO 2000/61739;WO 2001/29246;US2003/0115614;US2002/0164328;US2004/0093621;US 2004/0132140;US2004/0110704;US2004/0110282;US2004/0109865;WO 2003/085119;WO 2003/084570;WO 2005/035586;WO 2005/035778;WO 2005/053742;WO 2002/031140;Okazaki et al.J.Mol.Biol.336:1239-1249(2004);Yamane-Ohnuki et al.Biotech.Bioeng.87:614(2004). cell lines capable of producing defucosylated antibodies including Lec13 CHO cells lacking the fucosylation function of the protein (Ripka et al. Arch. Biochem. Biophys.249:533-545 (1986); US Pat Appl No US2003/0157108 A1,Presta,L; and WO 2004/056312A1,Adams et al, especially example 11), and knockout cell lines such as alpha-1, 6-fucosyltransferase genes, FUT8 knockout CHO cells (see Yamane-Ohnuki et al.Biotech.Bioeng.87:614(2004);Kanda,Y.et al.,Biotechnol.Bioeng.,94(4):680-688(2006); and WO 2003/085107).

Variants of anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) further provide bisecting oligosaccharides, e.g., wherein a double antennary oligosaccharide linked to the Fc region of an anti-TRAIL antibody is bisected by GlcNAc. Such anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) variants may have reduced fucosylation and/or enhanced ADCC function. Examples of such antibody variants are described in WO 2003/011878 (Jean-mair et al); U.S. Pat. No.6,602,684 (Umana et al); US 2005/0123346 (Umana et al), and FERRARA ET al, biotechnology and Bioengineering,93 (5): 851-861 (2006). Also provided are variants of anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) having at least one galactose residue in the oligosaccharide linked to the Fc region. Such anti-TRAIL antibody variants may have enhanced CDC function. Such variants are described, for example, in WO 1997/30087 (Patel et al); WO 1998/58964 (Raju, s.); and WO 1999/22764 (Raju, S.).

In some embodiments, the anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) variant comprises an Fc region capable of binding to fcyriii. In some embodiments, the anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) variant comprising an Fc region has ADCC activity in the presence of human effector cells (e.g., T cells) or has enhanced ADCC activity in the presence of human effector cells as compared to an otherwise identical anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) having a human wild-type IgG1 Fc region.

Cysteine engineered variants

In some embodiments, it is desirable to prepare cysteine engineered anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) in which one or more amino acid residues are substituted with cysteine residues. In some embodiments, the substitution residue occurs at an accessible site of the anti-TRAIL antibody. By substituting those residues with cysteines, active sulfhydryl groups located at accessible sites of anti-TRAIL antibodies can be used to couple the anti-TRAIL antibodies with other moieties, such as drug moieties or linker-drug moieties, to prepare anti-TRAIL immunoconjugates as further described herein. Cysteine engineered anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) may be prepared as described, for example, in u.s.pat.no.7,521,541.

Derivatives and their use as inhibitors of viral infection

In some embodiments, an anti-TRAIL antibody provided herein (e.g., a full length anti-TRAIL antibody) may be further modified to include other non-protein moieties known and readily available in the art. Suitable moieties for derivatizing anti-TRAIL antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol/propylene glycol copolymers, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1, 3-dioxolane, poly-1, 3, 6-trioxane, ethylene/maleic anhydride copolymers, polyaminoacids (homo-or random copolymers), dextran or poly (n-vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymers, propylene oxide/ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde has advantages in manufacturing due to its stability in water. The polymer may have any molecular weight and may be branched or unbranched. The number of polymers attached to the anti-TRAIL antibody may vary, and if more than one polymer is attached, they may be the same or different molecules. In general, the amount and/or type of polymer used for derivatization may be determined based on considerations including, but not limited to, the need to improve the properties or function of the anti-TRAIL antibody, whether the anti-TRAIL antibody derivative is used for treatment under particular conditions, and the like.

Pharmaceutical composition

Also provided herein are compositions (e.g., pharmaceutical compositions, also referred to herein as formulations) comprising any one of the anti-TRAIL antibodies (e.g., full-length anti-TRAIL antibodies), nucleic acids encoding the antibodies, vectors comprising nucleic acids encoding the antibodies, or host cells comprising the nucleic acids or vectors described herein. In some embodiments, a pharmaceutical composition is provided comprising any of the anti-TRAIL antibodies described herein and a pharmaceutically acceptable carrier.

Suitable anti-TRAIL antibody formulations may be prepared by mixing anti-TRAIL antibodies of the desired purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition,Osol,A.Ed (1980)) to obtain lyophilized formulations or liquid formulations. Acceptable carriers, excipients, or stabilizers are non-toxic to the recipient at the dosages and concentrations employed, and include buffers such as: phosphates, citric acid, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzyl ammonium chloride, hexamethyl ammonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butanol or benzyl alcohol, alkyl p-hydroxybenzoates such as methyl or propyl p-hydroxybenzoate, catechol, resorcinol, cyclohexanol, 3-pentanol and m-cresol); a low molecular weight (less than 10 residues) polypeptide; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., zinc-protein complexes); and/or nonionic surfactants such as TWEEN ^TM,PLURONICS^TM or polyethylene glycol (PEG); exemplary formulations are as described in WO98/56418 and are expressly incorporated herein by reference. Lyophilized formulations suitable for subcutaneous administration are described in WO 97/04801. Such lyophilized formulations can be reconstituted into high protein concentration formulations by means of a suitable diluent and the reconstituted formulations can be administered to the individual to be treated herein by means of subcutaneous administration. Cationic liposomes or liposomes can be used to deliver the anti-TRAIL antibodies of the application to cells.

The formulations described herein may contain, in addition to an anti-TRAIL antibody (e.g., a full length anti-TRAIL antibody), one or more other active agents necessary to treat a particular disorder, preferably agents that are complementary in activity and do not adversely affect each other. For example, it may be desirable to further include an anti-tumor agent, growth inhibitor, cytotoxic agent, or chemotherapeutic agent in addition to the anti-TRAIL antibody. These molecules are present in combination in amounts effective for the intended purpose. The effective amount of the other substances will depend on the amount of anti-TRAIL antibody in the formulation, the type of disease or disorder or treatment, and other factors as described above. These drugs are typically used at the same dosages and routes of administration as described herein, or at 1% to 99% of the presently employed dosages.

The anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) may also be embedded in microcapsules prepared, for example, by coacervation techniques and interfacial polymerization, such as hydroxymethylcellulose or gelatin-microcapsules and poly (methyl methacrylate) microcapsules, respectively, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Can be prepared into sustained release preparation.

Sustained release formulations of anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody (or fragments thereof), which matrices are in the form of shaped articles, e.g., films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly (2-hydroxyethyl methacrylate) or poly (vinyl alcohol)), polylactic acid (U.S. Pat. No.3,773,919), L-glutamic acid and L-ethyl glutamate copolymers, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON deptatm (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprorelin acetate), and poly-D (-) -3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic-glycolic acid can allow release of molecules for more than 100 days, certain hydrogels can release proteins in a shorter time. When encapsulated antibodies stay in the body for a long period of time, they may denature or aggregate as a result of exposure to a humid environment at 37 ℃ and may result in loss of biological activity or altered immunogenicity. The anti-TRAIL antibody can be stabilized according to a corresponding mechanism and a reasonable strategy. For example, if the aggregation mechanism is found to be the formation of intermolecular S-S bonds through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing in acidic solutions, controlling water content, using appropriate additives, and developing specific polymer matrix compositions.

In some embodiments, the anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) is formulated in a buffer containing citrate, sodium chloride, acetate, succinate, glycine, polysorbate 80 (tween 80), or any combination thereof.

Formulations for in vivo administration must be sterile. This can be easily achieved by, for example, filtration using sterile filtration membranes. Methods of treatment using anti-TRAIL antibodies

Anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) and/or compositions of the application may be administered to an individual (e.g., mammal, such as a human) to treat diseases and/or conditions associated with TRAIL signaling pathway (e.g., inflammatory diseases, autoimmune diseases, graft-related diseases, liver diseases, neurodegenerative diseases or cancers), including but not limited to transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic nephropathy, cancer. Accordingly, in some embodiments, the application provides a method of treating a disease and/or disorder associated with TRAIL signaling (e.g., an inflammatory disease, an autoimmune disease, a graft-related disease, a liver disease, a neurodegenerative disease, or cancer) comprising administering to an individual an effective amount of a composition (e.g., a pharmaceutical composition) comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), such as any of the anti-TRAIL antibodies described herein (e.g., a full-length anti-TRAIL antibody), in some embodiments, the individual is a human.

For example, in some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising a TRAIL antibody that specifically binds to an epitope on human TRAIL (e.g., a full-length anti-TRAIL antibody), wherein the epitope comprises the amino acid residues of human TRAIL. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 29, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 50 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 50; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 70 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 70.

In some embodiments, the anti-TRAIL antibodies described herein are full length anti-TRAIL antibodies comprising IgG1 or IgG4 constant regions. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 89. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 90. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 91. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO. 92.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 30, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 51 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 51; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 71 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 71.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 31, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 52 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 52; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 72 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 72.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising an amino acid sequence set forth in SEQ ID No. 53 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 53; and V _L, the V _L comprising the amino acid sequence shown in SEQ ID NO. 73 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown in SEQ ID NO. 73.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 54 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 54; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 74 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 74.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:20, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 55 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 55; and V _L, the V _L comprising an amino acid sequence set forth in SEQ ID No. 75 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 75.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 21, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 56 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 56; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 76 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 76.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 22, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 42, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 57 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 57; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 74 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 74.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 33, LC-CDR2 comprising the amino acid sequence SEQ ID No. 39, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 43, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 58 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 58; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 77 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 77.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 7, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO 34, LC-CDR2 comprising the amino acid sequence SEQ ID NO 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 59 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 59; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 78 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 78.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 60 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 60; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 79 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 79.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 61 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 61; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID No. 80 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 80.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 62 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 62; and V _L, the V _L comprising the amino acid sequence shown as SEQ ID NO. 81 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown as SEQ ID NO. 81.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:35, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 63 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 63; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 82 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 82.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 45, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 64 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 64; and V _L, the V _L comprising the amino acid sequence shown as SEQ ID NO. 83 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown as SEQ ID NO. 83.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 46, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 65 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 65; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 84 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 84.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 26, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:36, LC-CDR2 comprising the amino acid sequence SEQ ID NO:40, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:47, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 66 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 66; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 85 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 85.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 48, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 67 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 67; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 86 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 86.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 27, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO. 32, LC-CDR2 comprising the amino acid sequence SEQ ID NO. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO. 44, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 68 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 68; and V _L, the V _L comprising the amino acid sequence set forth in SEQ ID NO. 87 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID NO. 87.

For example, in some embodiments, there is provided a method for treating an individual having a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody), wherein the antibody comprises: a heavy chain variable domain (V _H), the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and a light chain variable domain (V _L), the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:49, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-TRAIL antibody is a full length antibody. In some embodiments, the full-length anti-TRAIL antibody is an IgG1 or IgG4 antibody. In some embodiments, the disease or condition is selected from, for example, transplant rejection, graft-versus-host disease, liver injury, pulmonary arterial hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer. In some embodiments, the individual is a human.

In some embodiments, a method is provided for treating an individual having a disorder associated with a TRAIL signaling pathway (e.g., an inflammatory disorder, an autoimmune disorder, a transplantation-related disorder, a liver disorder, a neurodegenerative disorder, or cancer), comprising administering to the individual an effective amount of a composition comprising an anti-TRAIL antibody, wherein the antibody comprises: v _H, said V _H comprising the amino acid sequence set forth in SEQ ID No. 69 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 69; and V _L, the V _L comprising the amino acid sequence shown in SEQ ID NO. 88 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown in SEQ ID NO. 88.

In some embodiments, the individual is a mammal (e.g., human, non-human primate, rat, mouse, cow, horse, pig, sheep, goat, dog, cat, etc.). In some embodiments, the individual is a human. In some embodiments, the individual is a clinical patient, a clinical trial volunteer, a laboratory animal, or the like. In some embodiments, the individual is less than 60 years old (including, for example, less than 50, 40, 30, 25, 20, 15, or 10 years old). In some embodiments, the individual is older than 60 years (including, for example, older than 70, 80, 90, or 100 years). In some embodiments, the individual is diagnosed with or genetically predisposed to one or more of the diseases or disorders described herein (e.g., inflammatory disease, autoimmune disease, graft-related disease, liver disease, neurodegenerative disease, or cancer). In some embodiments, the individual has one or more risk factors associated with one or more diseases or disorders described herein.

In some embodiments, the application provides a method of delivering an anti-TRAIL antibody (e.g., any of the anti-TRAIL antibodies described herein, e.g., an isolated anti-TRAIL antibody) to a cell expressing TRAIL on its surface in an individual, the method comprising administering to the individual a composition comprising an anti-TRAIL antibody.

Many diagnostic methods for inflammatory diseases, autoimmune diseases, transplantation-related diseases, liver diseases, neurodegenerative diseases or cancers or any other diseases that exhibit abnormal expression of TRAIL and clinical descriptions of these diseases are known in the art. Such methods include, but are not limited to, for example, immunohistochemistry, PCR, and Fluorescence In Situ Hybridization (FISH).

In some embodiments, the anti-TRAIL antibodies (e.g., full length anti-TRAIL antibodies) and/or compositions of the application are used in combination with a second, third, or fourth agent (including, e.g., an immunosuppressant, an anti-inflammatory agent, an antineoplastic agent, a growth inhibitor, a cytotoxic agent, a chemotherapeutic agent, or a vascular inhibitor) to treat a disease associated with TRAIL signaling pathway.

Dosage and method of administration of anti-TRAIL antibodies

The dosage of an anti-TRAIL antibody (e.g., an isolated anti-TRAIL antibody) composition administered to an individual (e.g., a human) may vary depending on the particular composition, mode of administration, and type of disease being treated. In some embodiments, the amount of the composition (e.g., a composition comprising an anti-TRAIL antibody) is effective to produce an objective response (e.g., a partial response or a complete response) in an inflammatory disease, an autoimmune disease, a transplant-related disease, a liver disease, a neurodegenerative disease, or a cancer treatment. In some embodiments, the amount of the anti-TRAIL antibody composition is sufficient to produce a complete response in the individual. In some embodiments, the amount of the anti-TRAIL antibody composition is sufficient to produce a partial response in the individual. In some embodiments, the amount of the anti-TRAIL antibody composition administered (e.g., when administered alone) is sufficient to produce a total response rate of greater than 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90% in a population of individuals treated with the anti-TRAIL antibody composition.

In some embodiments, the amount of the composition (e.g., a composition comprising an isolated anti-TRAIL antibody) is sufficient to extend the progression free survival of the individual. In some embodiments, the amount of the composition is sufficient to extend the overall survival of the individual. In some embodiments, the amount of the composition (e.g., when administered alone) is sufficient to produce a clinical benefit of greater than 50%, 60%, 70%, or 77% in a population of individuals treated with the anti-TRAIL antibody composition.

In some embodiments, the amount of a composition (e.g., a composition comprising an isolated anti-TRAIL antibody), alone or in combination with a second, third, and/or fourth agent, is an amount sufficient to control symptoms and reduce the risk of exacerbations prior to treatment or as compared to the corresponding activity in other subjects not receiving treatment. The magnitude of the therapeutic effect can be measured using standard methods, such as in vitro assays for purified enzymes, cell-based assays, animal models, or human trials.

In some embodiments, the amount of anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) in the composition is below a level that causes a toxic effect (i.e., an effect above a clinically acceptable toxicity level) or is at a level where potential side effects can be controlled or tolerated when the composition is administered to an individual.

In some embodiments, the amount of the composition approaches the Maximum Tolerated Dose (MTD) of the composition following the same dosing regimen. In some embodiments, the amount of the composition is greater than 80%, 90%, 95% or 98% of the MTD.

In some embodiments, the amount of anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) in the composition is in the range of 0.001 μg to 1000 μg.

In any of the embodiments described above, the effective amount of anti-TRAIL antibody (e.g., full length anti-TRAIL antibody) in the composition is in the range of 0.1 μg/kg to 100mg/kg as calculated on a weight basis.

The anti-TRAIL antibody composition may be administered to a subject (e.g., a human) by a variety of routes including, for example, intravenous, intra-arterial, intraperitoneal, intrapulmonary, oral, inhalational, intravascular, intramuscular, intratracheal, subcutaneous, intraocular, intrathecal, mucosal or transdermal. In some embodiments, a slow release formulation of the composition is used. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered through an artery. In some embodiments, the composition is administered intraperitoneally. In some embodiments, the composition is administered intrahepatially. In some embodiments, the composition is administered by hepatic arterial infusion. In some embodiments, the composition is applied to a site remote from the first lesion.

Product and kit

In some embodiments of the application, an article of manufacture is provided that comprises a substance that is capable of being used to treat a disease associated with TRAIL signaling pathway (e.g., an inflammatory disease, autoimmune disease, graft-related disease, liver disease, neurodegenerative disease, or cancer), or to deliver an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody) to cells that express TRAIL on the surface. The article of manufacture may comprise a container and a label or package insert attached to or associated with the container. Suitable containers include, for example, bottles, vials, syringes, and the like. The container may be made of a variety of materials, such as glass or plastic. Typically, the container contains a composition effective to treat the diseases or conditions described herein and has a sterile port (e.g., the container may be an iv bag or a vial with a pierceable cap of a hypodermic injection needle). At least one active substance in the composition is the anti-TRAIL antibody. The label or package insert identifies the particular condition for which the composition may be used. The label or package insert further comprises instructions for administering the anti-TRAIL antibody composition to the patient. Articles of manufacture and kits comprising combination therapies are within the contemplation herein.

Package insert refers to instructions that are typically contained within the commercial package of therapeutic products, including indications, usage, dosage, administration, contraindications, and/or warning information regarding the use of such therapeutic products. In some embodiments, the package insert indicates that the composition can be used to treat a disorder associated with TRAIL signaling (e.g., an inflammatory disorder, an autoimmune disorder, a transplant-related disorder, a liver disorder, a neurodegenerative disorder, or cancer). In some embodiments, the package insert indicates that the composition may be used to treat a disease including transplant rejection, graft-versus-host disease, liver injury, pulmonary hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis B, acute viral hepatitis C, chronic hepatitis B, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer.

In addition, the article of manufacture may further comprise a second container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffer, grignard solution, or dextrose solution. Other materials may be included as desired from a commercial and user standpoint, including other buffers, diluents, filters, needles and syringes.

Also, kits useful for various purposes, such as for treating diseases associated with TRAIL signaling pathways (e.g., inflammatory diseases, autoimmune diseases, transplantation-related diseases, liver diseases, neurodegenerative diseases, or cancers), or for delivering anti-TRAIL antibodies (e.g., full-length anti-TRAIL antibodies) to cells that express TRAIL on their surface, optionally in combination with a formulation. Kits of the application include one or more containers comprising an anti-TRAIL antibody composition (or single dose form and/or article of manufacture), and in some embodiments, further comprising another agent (e.g., an agent described herein) and/or instructions for use consistent with any of the methods described herein. The kit may further comprise a description of the selection of suitable individuals for treatment. The instructions for use attached to the kits of the application are typically written instructions on labels or packaging instructions (e.g., paper sheets contained within the kit), and machine-readable instructions (e.g., instructions on a magnetic or optical storage disc) are also acceptable.

For example, in some embodiments, the kit comprises a composition comprising an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody). In some embodiments, the kit comprises: a) A composition comprising any one of the anti-TRAIL antibodies described herein, and b) at least one other agent in an effective amount that enhances the effect (e.g., therapeutic effect, detection effect) of the anti-TRAIL antibody. In some embodiments, the kit comprises: a) A composition comprising any one of the anti-TRAIL antibodies described herein, and b) instructions for administering the anti-TRAIL antibody composition to an individual for treating a disease associated with TRAIL signaling pathway (e.g., an inflammatory disease, autoimmune disease, graft-related disease, liver disease, neurodegenerative disease, or cancer). In some embodiments, the kit comprises: a) a composition comprising any one of the anti-TRAIL antibodies described herein, and b) at least one additional agent in an amount effective to enhance the effect (e.g., therapeutic effect, detection effect) of the anti-TRAIL antibody, and c) instructions for administering the anti-TRAIL antibody composition and additional agents to an individual for treating a disorder associated with TRAIL signaling pathway (e.g., inflammatory disorder, autoimmune disorder, transplant-related disorder, liver disorder, neurodegenerative disorder, or cancer). The anti-TRAIL antibody and other substances may be present in separate containers or in the same container. For example, the kit may comprise one specific composition or two or more compositions, wherein one composition comprises an anti-TRAIL antibody and the other composition comprises another agent.

In some embodiments, the kit comprises a nucleic acid (or a set of nucleic acids) encoding an anti-TRAIL antibody (e.g., a full-length anti-TRAIL antibody). In some embodiments, the kit comprises: a) A nucleic acid (or a set of nucleic acids) encoding an anti-TRAIL antibody (e.g., a full length anti-TRAIL antibody), and b) a host cell expressing the nucleic acid (or the set of nucleic acids). In some embodiments, the kit comprises: a) A nucleic acid (or set of nucleic acids) encoding an anti-TRAIL antibody (e.g. a full length anti-TRAIL antibody), and b) instructions for use, adapted to: i) Expressing an anti-TRAIL antibody in a host cell, ii) preparing a composition comprising an anti-TRAIL antibody, and iii) administering the composition comprising an anti-TRAIL antibody to an individual to treat a disease associated with TRAIL signaling pathway (e.g., an inflammatory disease, autoimmune disease, transplantation-related disease, liver disease, neurodegenerative disease, or cancer). In some embodiments, the kit comprises: a) a nucleic acid (or set of nucleic acids) encoding an anti-TRAIL antibody (e.g., a full length anti-TRAIL antibody), b) a host cell expressing the nucleic acid (or set of nucleic acids), and c) instructions for use, suitable for: i) Expressing an anti-TRAIL antibody in a host cell, ii) preparing a composition comprising an anti-TRAIL antibody, and iii) administering the composition comprising an anti-TRAIL antibody to an individual to treat a disease associated with TRAIL signaling pathway (e.g., an inflammatory disease, autoimmune disease, transplantation-related disease, liver disease, neurodegenerative disease, or cancer).

The kit of the application is packaged in a suitable form. Suitable packages include, but are not limited to, vials, bottles, jars, flexible packages (e.g., sealed mylar or plastic bags), and the like. The kit may optionally provide additional components, such as buffers and instructional information. Thus, the present application also provides articles, including vials, bottles, jars, flexible packages (e.g., sealed mylar or plastic bags), and the like.

Instructions for use of the anti-TRAIL antibody composition typically include information such as dosage, period of administration, route of administration, and the like. The container may be unit dose, large package (e.g., multi-dose package) or subunit dose. For example, a kit comprising a sufficient dose of an anti-TRAIL antibody as described herein (e.g., a full length anti-TRAIL antibody) is provided for long term effective treatment of an individual, e.g., one week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, or more. The kit may also comprise multiple unit doses of anti-TRAIL antibodies, pharmaceutical compositions, and instructions for use, and be packaged in amounts sufficient for storage and use in a pharmacy, e.g., a hospital pharmacy and a compound pharmacy.

Those skilled in the art will recognize several embodiments that are possible within the scope and spirit of the application. The application will now be described in more detail by reference to the following non-limiting examples. The following examples further illustrate the application but should not be construed as in any way limiting its scope.

Detailed Description

Example 1: preparation of antigens

Preparation of recombinant TRAIL proteins

CDNA encoding the extracellular region of human TRAIL (human TRAIL; hTRAIL; genBank ID number: 8743) (synthesized by Beijing Yiqiao Shenzhou technologies Co., ltd.) was constructed into eukaryotic expression vectors by subcloning. His tag and/or other tags commonly used by those skilled in the art are added to the end of cDNA encoding the above protein, and a plasmid containing the human TRAIL extracellular region coding sequence is constructed. The above plasmid is transfected into 293F cells for expression to produce fusion proteins containing the TRAIL extracellular region, such as His-hTRAIL, where "His" represents a His tag.

The recombinant protein with His tag was purified by Immobilized Metal Affinity Chromatography (IMAC) using nickel (Ni) column-HISCAP SMART ff 5ml pre-packed column (SA 036C15, of zenithal and biotechnology limited) according to manufacturer's instructions. The specific operation is as follows: first, the nickel column was equilibrated with buffer A1 (50 mM Na ₃PO₄, 0.15M NaCl, pH 7.2) at a flow rate of 150cm/h, and the pH of the solution containing the fusion protein (e.g., cell culture supernatant) was adjusted to 7.2, and the sample was loaded at room temperature at a flow rate of 150cm/h. Subsequently, the column was again equilibrated with 6 column volumes of A1 buffer at a flow rate of 150cm/h. Finally, elution was performed with 10 column volumes of 50mM PB solution (containing 0.15M NaCl and 0.2M imidazole, pH 7.2) and the eluate was collected. Ultrafiltering the obtained solution into PBS by using an ultrafilter tube, and concentrating.

Example 2: screening for Single chain antibodies (scFv) against TRAIL

2.1 Construction of scFv antibody phage display library:

Mice were immunized with His-htril as antigen together with an equal volume (v/v) of adjuvant. Serum from immunized mice was taken and tested for total IgG titers by ELISA. After several rounds of immunization, phage display libraries were established using the spleen of the mice. Briefly, the spleen of immunized mice was taken, RNA was extracted, cDNA was obtained by reverse transcription, V _H and V _K fragments were amplified using V _H and V _K specific primers, and after gel recovery purification, V _H and V _K were ligated to construct scFv, which was cloned into phage display plasmids, which were then electrotransferred into e.coli TG1, and phage infected with e.coli TG1 to obtain scFv antibody phage display libraries.

2.2 Screening for anti-TRAIL single chain antibodies (scFv):

scFv that specifically bound human TRAIL were isolated and obtained from phage display libraries after multiple rounds of panning. Briefly, a 2X10 ¹¹ PFU phage scFv library was added to His-hTRAIL antigen coated ELISA plates and incubated at 37℃for 2 hours. The phage bound to human TRAIL antigen was captured by TRAIL antigen coated on ELISA plates and unbound phage was washed off using PBST solution for 8-15 washes (wash times increased with increasing number of screening rounds). Phage that specifically bound to hTRAIL antigen were eluted with 0.1M Glycine-HCl solution (ph 2.2). The phage was infected with TG1 cells in exponential growth phase, and after 1 hour of culture with ampicillin, helper phage was added thereto, and the culture was performed overnight at 28℃and 220rpm in a shaker. The culture solution is collected the next day, supernatant is obtained after centrifugation, and the next round of screening is performed. Positive scFv antibody libraries were obtained after multiple rounds of panning.

2.3ELISA method for detection of anti-TRAIL Single-chain antibodies:

ELISA screening was performed on phages enriched after panning. Briefly, his-hTRAIL antigen was dissolved in PBS solution and coated in 96-well plates at 0.1. Mu.g/well overnight at 4 ℃. After washing 1 time with PBST solution, 90 μl of PBS containing 4% nonfat dry milk was added to each well. Subsequently, 10. Mu.L of the supernatant of the phase-scFv culture was added to the corresponding wells and incubated at 37℃for 1-2 hours. After washing 8 times with PBST solution, 100. Mu.L/well of anti-M13-HRP antibody (Sino Biological,11973-MM 05T-H) was added at a 1:4000 dilution. Incubate at 37℃for 1h. After washing the plates 6 times with PBST, TMB color development solution was added, 100. Mu.L/well, and incubated at room temperature for 10min in the dark. The chromogenic reaction was stopped with 2M H ₂SO₄ and the absorbance at 450nm was read with a microplate reader. At the end of the screening process, a series of positive scFv antibodies were obtained and sequenced.

Example 3: preparation and characterization of full-Length chimeric anti-TRAIL antibodies

3.1 Preparation of full Length anti-TRAIL chimeric antibodies

The obtained positive scFv antibody was reconstituted into a chimeric antibody having a human IgG1 or IgG4 heavy chain constant region and a human kappa (κ) light chain constant region. V _L and V _H were amplified from phage display vectors and constructed into eukaryotic expression vectors pTTa-L (containing human kappa constant region) and pTT5-H1 (containing human IgG1 heavy chain constant region) or pTTa-H4 (containing human IgG4 heavy chain constant region), respectively. The 293F cells were CO-transfected with the plasmid expressing the light chain and the plasmid expressing the heavy chain, and cultured at 37℃with 5% CO ₂ at 120rpm for 6 days, and the culture broth was purified using protein A column-MabCap At FF 5ml pre-packed column (Hemsleyakulare and Biotech Co., ltd., SA023C 15). The specific operation is as follows: the protein A column was first equilibrated with 6 column volumes of PBS buffer (containing 50mM PB and 0.15M NaCl, pH 7.2) at a flow rate of 5ml/min. The pH of the culture supernatant was adjusted to 7.2, and the sample was applied at room temperature at a flow rate of 5ml/min. Subsequently, the column was equilibrated again with 6-10 column volumes of PBS buffer at a flow rate of 5ml/min. After complete equilibration, the eluate (containing 0.1M Gly and 150mM NaCl, pH 3.2) was added for elution and the eluate was collected. Ultrafiltering the obtained solution into PBS by using an ultrafilter tube, and concentrating. After ultrafiltration concentration, the antibody concentration was determined and further biochemical and biological activity assays were performed.

3.2 Activity assay of anti-TRAIL antibodies

Anti-TRAIL antibodies inhibit TRAIL antigen-induced apoptosis of HepG2 cells:

Exogenous TRAIL activates the apoptosis signaling pathway and mediates apoptosis in the case of inhibiting cell division with the addition of actinomycin D. The ATP content of a cell may reflect the viability of the cell, inversely proportional to the degree of apoptosis. Based on the above principle, can be utilized The cell viability detection kit by the luminescence method detects the ATP content, and further determines the activity of the anti-TRAIL chimeric antibody (reconstructed into a human IgG4 form) for inhibiting the apoptosis of the HepG2 cell induced by the TRAIL antigen.

HepG2 cells in the logarithmic growth phase (purchased from the cell bank of the Beijing institute of synergetic, 1101HUM-PUMC 000035) were seeded in 96-well plates, 2X 10 ⁴ cells/well. Each antibody sample was diluted to 108nM and subsequently diluted in a specific ratio. 50. Mu.L of actinomycin D (Hanhui pharmaceutical Co., ltd., national drug standard H20023504), 25. Mu.L of TRAIL recombinant fusion protein (Novoprotein, C022) and 25. Mu.L of diluted anti-TRAIL antibody were added to a 96-well plate containing HepG2 cells so that the final concentration of actinomycin D, TRAIL recombinant fusion protein was 1. Mu.g/mL and 0.5ng/mL, respectively. The cells were incubated at 37℃for 48 hours under 5% CO ₂. By means ofThe luminescence cell viability assay kit (Promega, G7572) detects the ATP content, which is proportional to the luminescence signal generated. Namely, adding to 96-well cell culture platesReagents, 50 μl per well, were mixed and incubated at room temperature for 10min. The fluorescence value was read with a microplate reader. Curves were generated by GRAPHPAD PRISM software and IC ₅₀ values for each anti-TRAIL antibody were calculated.

The results are shown in Table 5, the anti-TRAIL chimeric antibodies H12, H13, H15, H16, H17, H18, H19, H20, H27, H48, H50, H51, H52, H53, H55, H62, H63 and H71 all have obvious inhibition effect on the apoptosis of HepG2 cells induced by TRAIL antigen, and the ability of the anti-TRAIL chimeric antibodies to inhibit the apoptosis of HepG2 cells induced by TRAIL antigen is obviously superior to or equivalent to that of the control MAB375-500 (commercial anti-TRAIL antibody, RD).

Table 5: anti-TRAIL chimeric antibodies inhibiting TRAIL antigen-induced apoptosis of HepG2 cells

Antibody name	IC₅₀(nM)	Antibody name	IC₅₀(nM)
				H12	0.07882	H50	0.1084
H13	0.02293	H51	0.0555
				H15	0.05273	H52	0.05696
H16	0.02032	H53	0.07913
				H17	0.1003	H55	0.1238
H18	0.008689	H62	0.07623
				H19	0.01816	H63	0.1045
H20	0.03059	H71	0.1527
				H27	0.09537	MAB375-500	0.1027
H48	0.06393

3.3 Determination of binding Capacity of anti-TRAIL antibody

The constructed full length anti-TRAIL chimeric antibody (reconstituted to human IgG4 form) was subjected to a binding assay with human TRAIL to reflect the binding activity of the anti-TRAIL antibody to human TRAIL antigen. Briefly, his-hTRAIL antigen was dissolved in PBS solution and coated in 96-well plates at 0.1. Mu.g/well overnight at 4 ℃. The mixture was blocked with 5% milk at 37℃for 1 hour and washed 6 times with PBST solution. Each antibody sample was first diluted to 133nM, followed by 1:4, and carrying out gradient dilution according to the proportion. Samples after gradient dilution were added to the coated 96-well plates, 100. Mu.L per well, and incubated at 37℃for 1 hour. Followed by 6 washes with PBST solution. To each well 100. Mu.L of anti-human IgG-HRP (Sigma, A8667,1:10000 dilution) was added and incubated for 1 hour at 37 ℃. Wash 6 times with PBST solution. 100. Mu.L TMB was added to each well and incubated at 37℃for 10-20 minutes, and the reaction was stopped with 2M H ₂SO₄. The absorbance at 450nm was read using an microplate reader. Binding curves were generated by GRAPHPAD PRISM and EC ₅₀ values were calculated for each anti-TRAIL antibody.

As a result, as shown in Table 6, the anti-TRAIL chimeric antibodies H12, H13, H15, H16, H17, H18, H19, H20, H27, H48, H50, H51, H52, H53, H55, H62, H63, H71 were selected to have good binding ability to human TRAIL antigen, and the binding ability was comparable to that of the control antibody MAB 375-500.

Table 6: binding ability of anti-TRAIL chimeric antibodies to human TRAIL

Antibody name	EC₅₀(nM)	Antibody name	EC₅₀(nM)
				H12	0.285	H51	0.4261
H13	0.3159	H52	0.4283
				H15	0.3349	H53	0.6058
H16	0.4641	H55	0.6233
				H17	0.6581	H62	0.7315
H18	0.2106	H63	0.7113
				H19	0.4228	H71	0.4024
H20	0.3571	H73	0.2498
				H27	0.3094	H76	0.2625
H48	0.7302	MAB375-500	0.27
				H50	0.3671

Claims

1. An isolated antibody that specifically recognizes TRAIL, wherein the anti-TRAIL antibody comprises:

(i) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 50V _H;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 70;

(ii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 51V _H;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 71;

(iii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 52V _H;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 72;

(iv) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 53V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 73;

(v) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 54V _H;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 74;

(vi) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 55V _H;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 75;

(vii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 56V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 76;

(viii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 57V _H;

(ix) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 58V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 77;

(x) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID NO 59V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 78;

(xi) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 60V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 79;

(xii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 61V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 80;

(xiii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 62V _H;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 81;

(xiv) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 63V _H;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 82;

(xv) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 64V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 of V _L as shown in amino acid sequence SEQ ID NO 83;

(xvi) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 65V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 84;

(xvii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 66, which V _H comprises;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO: 85;

(xviii) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by the amino acid sequence SEQ ID No. 67V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 86;

(xix) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by V _H as shown in amino acid sequence SEQ ID NO 68;

and V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 87; or (b)

(Xx) V _H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as represented by amino acid sequence SEQ ID No. 69V _H;

And V _L comprising the LC-CDR1, LC-CDR2 and LC-CDR3 comprised by V _L as shown in amino acid sequence SEQ ID NO 88.

2. An isolated antibody that specifically recognizes TRAIL, wherein the antibody comprises:

(i) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 29, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(ii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 30, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(iii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 2, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 18, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 31, LC-CDR2 comprising the amino acid sequence SEQ ID No. 37, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 41, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(iv) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(v) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 19, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(vi) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:20, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(vii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 5, HC-CDR2 comprising the amino acid sequence SEQ ID NO 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 21, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(viii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 4, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 12, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 22, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 42, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(ix) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 6, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 13, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 23, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 33, LC-CDR2 comprising the amino acid sequence SEQ ID No. 39, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 43, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(x) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 7, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 34, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 44, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xi) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 44, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 45, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xiii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 35, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 45, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xiv) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 9, HC-CDR2 comprising the amino acid sequence SEQ ID NO 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 45, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xv) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 25, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 46, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xvi) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO 10, HC-CDR2 comprising the amino acid sequence SEQ ID NO 16, and HC-CDR3 comprising the amino acid sequence SEQ ID NO 26, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 36, LC-CDR2 comprising the amino acid sequence SEQ ID No. 40, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 47, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xvii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 14, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 24, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 48, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs;

(xviii) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 8, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 15, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 27, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID No. 32, LC-CDR2 comprising the amino acid sequence SEQ ID No. 38, and LC-CDR3 comprising the amino acid sequence SEQ ID No. 44, or a variant of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs; or (b)

(Xix) V _H, the V _H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO. 11, HC-CDR2 comprising the amino acid sequence SEQ ID NO. 17, and HC-CDR3 comprising the amino acid sequence SEQ ID NO. 28, or variants of said V _H comprising up to about 5 amino acid substitutions in the HC-CDRs; and V _L, the V _L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:38, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:49, or variants of said V _L comprising up to about 5 amino acid substitutions in the LC-CDRs.

3. An isolated antibody specifically recognizing TRAIL according to claim 1 or 2, which comprises:

(i) V _H comprising the amino acid sequence set forth in SEQ ID No. 50 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 50; and V _L comprising the amino acid sequence set forth in SEQ ID No. 70 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 70;

(ii) V _H comprising the amino acid sequence set forth in SEQ ID No. 51 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 51; and V _L comprising the amino acid sequence set forth in SEQ ID No. 71 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 71;

(iii) V _H comprising the amino acid sequence set forth in SEQ ID No. 52 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 52; and V _L comprising the amino acid sequence set forth in SEQ ID No. 72 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 72;

(iv) V _H comprising the amino acid sequence set forth in SEQ ID No. 53 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 53; and V _L comprising the amino acid sequence set forth in SEQ ID No. 73 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 73;

(v) V _H comprising the amino acid sequence set forth in SEQ ID No. 54 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 54; and V _L comprising the amino acid sequence set forth in SEQ ID No. 74 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 74;

(vi) V _H comprising the amino acid sequence set forth in SEQ ID No. 55 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 55; and V _L comprising the amino acid sequence set forth in SEQ ID No. 75 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 75;

(vii) V _H comprising the amino acid sequence set forth in SEQ ID No. 56 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 56; and V _L comprising the amino acid sequence set forth in SEQ ID No. 76 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 76;

(viii) V _H comprising the amino acid sequence set forth in SEQ ID No. 57 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 57; and V _L comprising the amino acid sequence set forth in SEQ ID No. 74 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 74;

(ix) V _H comprising the amino acid sequence set forth in SEQ ID No. 58 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 58; and V _L comprising the amino acid sequence set forth in SEQ ID No. 77 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 77;

(x) V _H comprising the amino acid sequence set forth in SEQ ID No. 59 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 59; and V _L comprising the amino acid sequence set forth in SEQ ID No. 78 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 78;

(xi) V _H comprising the amino acid sequence set forth in SEQ ID No. 60 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 60; and V _L comprising the amino acid sequence set forth in SEQ ID No. 79 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 79;

(xii) V _H comprising the amino acid sequence set forth in SEQ ID No. 61 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 61; and V _L comprising the amino acid sequence set forth in SEQ ID No. 80 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 80;

(xiii) V _H comprising the amino acid sequence set forth in SEQ ID No. 62 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 62; and V _L comprising the amino acid sequence shown as SEQ ID No. 81 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown as SEQ ID No. 81;

(xiv) V _H comprising the amino acid sequence set forth in SEQ ID No. 63 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 63; and V _L comprising the amino acid sequence set forth in SEQ ID No. 82 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 82;

(xv) V _H comprising the amino acid sequence set forth in SEQ ID No. 64 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 64; and V _L comprising the amino acid sequence shown as SEQ ID NO. 83 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown as SEQ ID NO. 83;

(xvi) V _H comprising the amino acid sequence set forth in SEQ ID No. 65 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 65; and V _L comprising the amino acid sequence set forth in SEQ ID No. 84 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 84;

(xvii) V _H comprising the amino acid sequence set forth in SEQ ID No. 66 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 66; and V _L comprising the amino acid sequence set forth in SEQ ID No. 85 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 85;

(xviii) V _H comprising the amino acid sequence set forth in SEQ ID No. 67 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 67; and V _L comprising the amino acid sequence set forth in SEQ ID No. 86 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 86;

(xix) V _H comprising the amino acid sequence set forth in SEQ ID No. 68 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 68; and V _L comprising the amino acid sequence set forth in SEQ ID No. 87 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 87; or (b)

(Xx) V _H comprising the amino acid sequence set forth in SEQ ID No. 69 or a variant thereof having at least about 80% sequence identity to the amino acid sequence set forth in SEQ ID No. 69; and V _L comprising the amino acid sequence shown as SEQ ID NO. 88 or a variant thereof having at least about 80% sequence identity to the amino acid sequence shown as SEQ ID NO. 88.

4. An isolated antibody that specifically recognizes TRAIL according to any one of claims 1-3, wherein the antibody comprises an Fc fragment.

5. The isolated antibody that specifically recognizes TRAIL according to claim 4, wherein the antibody is a full-length IgG antibody.

6. The isolated antibody that specifically recognizes TRAIL according to claim 5, wherein the antibody is a full length IgG1, igG2, igG3, or IgG4 antibody.

7. An antibody specifically recognizing TRAIL isolated according to any one of claims 1-6, wherein the antibody is a chimeric, humanized or fully human antibody.

8. An isolated antibody that specifically recognizes TRAIL according to any one of claims 1-3, which antigen binding fragment is selected from the group consisting of Fab, fab ', F (ab) ' ₂, fab ' -SH, single chain antibody (scFv), fv fragment, dAb, fd, nanobody (nanobody), diabody (diabody) and linear antibody.

9. A nucleic acid molecule encoding the antibody of any one of claims 1-8 that specifically recognizes TRAIL.

10. A vector comprising the nucleic acid molecule of claim 9.

11. An isolated host cell comprising an antibody of any one of claims 1-8 that specifically recognizes TRAIL, a nucleic acid molecule of claim 9, or a vector of claim 10.

12. A method of making an antibody that specifically recognizes TRAIL, comprising:

a) Culturing the host cell of claim 11 under conditions effective to express an antibody that specifically recognizes TRAIL; and

B) The expressed antibodies that specifically recognize TRAIL are obtained from the host cell.

13. A pharmaceutical composition comprising an antibody that specifically recognizes TRAIL according to any one of claims 1-8, a nucleic acid molecule according to claim 9, a vector according to claim 10, an isolated host cell according to claim 11, or an antibody produced by the method according to claim 12, and a pharmaceutically acceptable carrier.

14. Use of an antibody according to any one of claims 1-8, a nucleic acid molecule according to claim 9, a vector according to claim 10, an isolated host cell according to claim 11, an antibody produced by a method according to claim 12, or a pharmaceutical composition according to claim 13 in the manufacture of a medicament for the treatment of a disease or disorder in a subject in need thereof.

15. A method of treating a disease or disorder, the method comprising administering to an individual in need thereof an effective amount of an antibody of any one of claims 1-8, a nucleic acid molecule of claim 9, a vector of claim 10, an isolated host cell of claim 11, an antibody produced by the method of claim 12, or a pharmaceutical composition of claim 13.

16. The use according to claim 14 or the method of treatment according to claim 15, wherein the disease or condition is an inflammatory disease, autoimmune disease, transplantation related disease, liver disease, neurodegenerative disease or cancer associated with TRAIL signaling.

17. The use or method of treatment according to claim 16, wherein the disease or condition is selected from the group consisting of transplant rejection, graft-versus-host disease, liver injury, pulmonary hypertension, alzheimer's disease, acute lung injury, acute respiratory distress syndrome, myocardial infarction, cardiomyopathy, ischemic reperfusion injury, diabetes, brain injury, spinal cord injury, acute viral hepatitis b, acute viral hepatitis c, chronic hepatitis b, alcoholic hepatitis, non-alcoholic steatohepatitis, cirrhosis, drug-induced liver injury/liver failure, autoimmune hepatitis, chronic kidney disease, acute kidney disease, diabetic kidney disease, cancer.