Anti - tryptase antibody, its composition, and its use

Abstract

To provide compositions including anti-tryptase antibodies and pharmaceutical compositions thereof, as well as methods of using the same.SOLUTION: A pharmaceutical for treating a pulmonary disorder comprises an antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof. The antibody comprises the following six hypervariable regions (HVRs): (a) an HVR-H1 comprising the amino acid sequence of DYGMV; (b) an HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG; (c) an HVR-H3 comprising the amino acid sequence of RNYDDWYFDV; (d) an HVR-L1 comprising the amino acid sequence of SASSSVTYMY; (e) an HVR-L2 comprising the amino acid sequence of RTSDLAS; and (f) an HVR-L3 comprising the amino acid sequence of QHYHSYPLT.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 457,722, filed on February 10, 2017, which is hereby incorporated by reference in its entirety.

[0002] Sequence Listing This application is being filed electronically in ASCII format and includes a sequence listing that is hereby incorporated by reference in its entirety. The ASCII copy, created on February 9, 2018, is named 50474 - 112WO2_Sequence_Listing_2.9.18_ST25 and is 108,967 bytes in size.

[0003] The present invention relates to anti - tryptase antibodies, pharmaceutical compositions, and methods of using the same.

Background Art

[0004] Human tryptase beta is a trypsin - like serine protease that is abundant in mast cells and, to a lesser extent, in basophils. Human tryptase beta, produced by the TPSAB1 and TPSB2 loci (where its three subtypes, tryptase beta 1, tryptase beta 2, and tryptase beta 3 exist), is the dominant active tryptase produced by human mast cells. These two loci produce four tryptase isoforms, where TPSAB1 produces tryptase alpha and tryptase beta 1, while TPSB2 produces tryptase beta 2 and tryptase beta 3. Tryptase alpha, as well as other isoforms such as tryptase gamma, tryptase delta, and tryptase epsilon, are mostly inactive.

[0005] Active tryptase beta, which has been processed for proteolytic activity, is stored in the secretory granules of mast cells as a heparin-complexed tetramer. Degranulation of mast cells, which can be induced by IgE-dependent stimuli (e.g., allergens) or non-IgE-dependent stimuli (e.g., substance P or active tryptase), results in the release of tryptase beta along with other granule enzymes and histamine. Previous studies have observed increased numbers of mast cells in the bronchial smooth muscle and epithelium of asthmatic patients, as well as increased levels of tryptase beta in bronchoalveolar lavage fluid. In addition, tryptase has been shown to contribute to airway bronchoconstriction and airway hyperreactivity, and also to play a role in idiopathic pulmonary fibrosis and extracellular matrix turnover, which demonstrate the airway remodeling process.

[0006] Tryptase has been implicated in various diseases and disorders, including asthma and other lung disorders, inflammatory disorders, autoimmune deficiencies, and fibrotic disorders, for which there remains a need for improved treatments and methods of treatment, including therapeutic anti-tryptase antagonists. Attempts have been made to develop small molecule tryptase inhibitors (see, e.g., Cairns, J.A., 2005, Pulmonary Pharmacology & Therapeutics 18:55-66), however, to the best of our knowledge, biological tryptase antagonist therapies, particularly anti-tryptase antagonist antibodies, have not been reported. SUMMARY OF THE INVENTION

[0007] The present invention relates to anti-tryptase antibodies, pharmaceutical compositions thereof, and methods of using the same.

[0008] In one aspect, the invention features an isolated antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, wherein the antibody comprises the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8), (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some embodiments, the antibody is defined by the six HVRs comprising the amino acid sequences of SEQ ID NOs: 7, 2, 8, 4, 5, and 6. In some embodiments, the antibody further comprises S43, P46, and W47 (Kabat numbering) in the light chain variable (VL) domain framework region L2 (FR-L2). In some embodiments, the antibody comprises (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 9, (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 10, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody further comprises the following VH domain framework regions (FRs): (a) FR-H1 comprising the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFS (SEQ ID NO: 11), (b) FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 12), (c) FR-H3 comprising the amino acid sequence of RFTISRDNSKNTLYLQMNSLRAEDTAVYYCTR (SEQ ID NO: 13), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14). In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 9.In some embodiments, the antibody further comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 15), (b) FR-L2 comprising the amino acid sequence of WYQQKPGKSPKPWIY (SEQ ID NO: 16), (c) FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 17), and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 18). In some embodiments, the VL domain comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 77. In other embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 78 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 79.

[0009] In another aspect, the invention features an isolated antibody, or an antigen-binding fragment thereof, that binds to human tryptase beta 1, the antibody comprising (a) a VH domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 9, and (b) a VL domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 9 and a VL domain comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 77. In other embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 78 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 79.

[0010] In another aspect, the invention features an isolated antibody comprising (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 77.

[0011] In another aspect, the invention features an isolated antibody comprising (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 78 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 79.

[0012] In another aspect, the invention features an isolated antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, the antibody comprising the following six HVRS: (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RDNYDWYFDV (SEQ ID NO: 29), (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVKLVESGGGSVQPGGSRKLSCAASGFTFS (SEQ ID NO: 21), (b) FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 22), (c) FR-H3 comprising the amino acid sequence of RFTISRDNPKNTLFLQMSSLRSEDTAMYYCAR (SEQ ID NO: 23), and (d) FR-H4 comprising the amino acid sequence of WGTGTTVTVSS (SEQ ID NO: 24). In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, the antibody further comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of QIVLTQSPAIMSASPGEKVTISC (SEQ ID NO: 25), (b) FR-L2 comprising the amino acid sequence of WYQQKPGSSPKPWIY (SEQ ID NO: 26), (c) FR-L3 comprising the amino acid sequence of GVPARFSGSGSGTSYSLTISSMEAEDAATYYC (SEQ ID NO: 27), and (d) FR-L4 comprising the amino acid sequence of FGAGTKLELK (SEQ ID NO: 28). In some embodiments, the VL domain comprises the amino acid sequence of SEQ ID NO: 20.

[0013] In another aspect, the present invention features an isolated antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, wherein the antibody comprises (a) a VH domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 19, (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 20, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVKLVESGGGSVQPGGSRKLSCAASGFTFS (SEQ ID NO: 21), (b) FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 22), (c) FR-H3 comprising the amino acid sequence of RFTISRDNPKNTLFLQMSSLRSEDTAMYYCAR (SEQ ID NO: 23), and (d) FR-H4 comprising the amino acid sequence of WGTGTTVTVSS (SEQ ID NO: 24). In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, the antibody further comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of QIVLTQSPAIMSASPGEKVTISC (SEQ ID NO: 25), (b) FR-L2 comprising the amino acid sequence of WYQQKPGSSPKPWIY (SEQ ID NO: 26), (c) FR-L3 comprising the amino acid sequence of GVPARFSGSGSGTSYSLTISSMEAEDAATYYC (SEQ ID NO: 27), and (d) FR-L4 comprising the amino acid sequence of FGAGTKLELK (SEQ ID NO: 28). In some embodiments, the VL domain comprises the amino acid sequence of SEQ ID NO: 20.

[0014] In another aspect, the present invention features an isolated antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, comprising (a) a VH domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 19, and (b) a VL domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 20.

[0015] In some embodiments of any of the foregoing aspects, the antibody binds to an epitope on human tryptase beta 1 that comprises at least 1, at least 2, at least 3, or all 4 residues selected from the group consisting of His51, Val80, Lys81, and Asp82 of SEQ ID NO: 71. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 that comprises at least 1, at least 2, at least 3, or all 4 residues selected from the group consisting of His51, Val80, Lys81, and Asp82 of SEQ ID NO: 71. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 that comprises His51 of SEQ ID NO: 71 and at least 1, at least 2, or all 3 residues selected from the group consisting of Val80, Lys81, and Asp82. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or all 12 amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises His51, Gln67, Val80, Lys81, Asp82, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope is associated with the human tryptase beta 1 monomer or tetramer. In some embodiments, the epitope is determined by an X-ray crystallography model.In some embodiments, the antibody is capable of dissociating both the small interface of tetrameric human tryptase beta1 and the large interface of tetrameric human tryptase beta1.

[0016] In another aspect, the invention features an isolated antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, wherein the antibody comprises the following six HVRS: (a) HVR-H1 comprising the amino acid sequence of GYAIT (SEQ ID NO: 30), (b) HVR-H2 comprising the amino acid sequence of GISSAATTFYSSWAKS (SEQ ID NO: 31), (c) HVR-H3 comprising the amino acid sequence of DPRGYGAALDRLDL (SEQ ID NO: 32), (d) HVR-L1 comprising the amino acid sequence of QSIKSVYNNRLG (SEQ ID NO: 33), (e) HVR-L2 comprising the amino acid sequence of ETSILTS (SEQ ID NO: 34), and (f) HVR-L3 comprising the amino acid sequence of AGGFDRSGDTT (SEQ ID NO: 35). In some embodiments, the antibody is defined by the six HVRS comprising the amino acid sequences of SEQ ID NOs: 30, 31, 32, 33, 34, and 35. In some embodiments, the antibody further comprises Arg71 and Val78 (Kabat numbering) in VH domain FR-H3. In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVQLVESGPGLVKPSETLSLTCTVSRFSLI (SEQ ID NO: 38), (b) FR-H2 comprising the amino acid sequence of WIRQPPGKGLEWIG (SEQ ID NO: 42), (c) FR-H3 comprising the amino acid sequence of RVTISRDTSKNQVSLKLSSVTAADTAVYYCAR (SEQ ID NO: 43), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41). In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 36. In some embodiments, the antibody comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 64), (b) FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 65), (c) FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 66), and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 63). In some embodiments, the VL domain comprises the amino acid sequence of SEQ ID NO: 37.In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 81. In other embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 83.

[0017] In another aspect, the present invention features an isolated antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, wherein the antibody comprises (a) a VH domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 36, 47, 48, 49, 50, 51, and 52, (b) a VL domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% identity with any one of the amino acid sequences of SEQ ID NOs: 37, 53, 58, or 59, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVQLVESGPGLVKPSETLSLTCTVSRFSLI (SEQ ID NO: 38), (b) FR-H2 comprising the amino acid sequence of WIRQPPGKGLEWIG (SEQ ID NO: 42), (c) FR-H3 comprising the amino acid sequence of RVTISRDTSKNQVSLKLSSVTAADTAVYYCAR (SEQ ID NO: 43), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41). In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 36. In some embodiments, the antibody comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 64), (b) FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 65), (c) FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 66), and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 63). In some embodiments, the VL domain comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 81. In other embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 83.

[0018] In another aspect, the present invention features an isolated antibody that binds to human tryptase, or an antigen-binding fragment thereof, wherein the antibody comprises (a) a VH domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 36, and (b) a VL domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 37. In some embodiments, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 36 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 81. In other embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 83.

[0019] In another aspect, the present invention features an isolated antibody comprising (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 81.

[0020] In another aspect, the present invention features an isolated antibody comprising (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 83.

[0021] In another aspect, the present invention features an isolated antibody that binds to human tryptase, or an antigen-binding fragment thereof, wherein the antibody comprises (a) a VH domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 52, and (b) a VL domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 53.

[0022] In some embodiments of any of the foregoing aspects, the antibody binds to an epitope on human tryptase beta 1 comprising at least 1, at least 2, or all 3 residues selected from the group consisting of Gln100, Leu101, and Leu102 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or all 14 amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71. In some embodiments, the epitope comprises Gln35, Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Gln100, Leu101, Leu102, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, Glu129, and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is associated with the human tryptase beta 1 monomer or tetramer. In some embodiments, the epitope is associated with the human tryptase beta 1 tetramer and the epitope on human tryptase beta 1 further comprises one or both of Gln35 and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is determined by an X-ray crystallography model. In some embodiments, the antibody is capable of dissociating the small interface and / or the large interface of human tryptase beta 1.

[0023] In some embodiments of any of the foregoing aspects, the antibody further binds to cynomolgus (cyno) tryptase. In some embodiments, the antibody further binds to human tryptase alpha. In some embodiments, the antibody further binds to human tryptase beta2 or human tryptase beta3. In some embodiments, the antibody binds to both human tryptase beta2 and human tryptase beta3.

[0024] In some embodiments of any of the foregoing aspects, the antibody binds to tryptase with a K D of about 1 nM or less. In some embodiments, the K D is measured by surface plasmon resonance (SPR) assay. In some embodiments, the antibody binds to tryptase with a K D of about 120 pM to about 0.5 nM. In some embodiments, the antibody binds to tryptase with a K D of about 120 pM to about 300 pM. In some embodiments, the antibody binds to tryptase with a K D of about 120 pM to about 200 pM. In some embodiments, the antibody binds to tryptase with a K D of about 180 pM. In some embodiments, the antibody binds to tryptase with a K D of about 400 pM. In some embodiments, the SPR assay is performed at 25°C. In some embodiments, the K Dis measured, for example, using a BIACORE® SPR assay as described in section (A)(vii) of Example 1. In some embodiments, the SPR assay can use a BIAcore® T200 or equivalent instrument. In some embodiments, a BIAcore® Series S CM5 sensor chip (or equivalent sensor chip) is immobilized with a monoclonal mouse anti-human IgG (Fc) antibody, and the anti-tryptase antibody is sequentially captured on the flow cell. Serial three-fold dilutions of His-tagged human tryptase beta 1 monomer (SEQ ID NO: 128) are injected at a flow rate of 30 μl / min. Each sample is analyzed for 3 minutes association and 10 minutes dissociation. The assay is performed at 25°C. After each injection, the chip is regenerated using 3 M MgCl2. The binding response is corrected by subtracting the response units (RU) from a flow cell capturing an unrelated IgG of similar density. k on and k off of the simultaneous fitting of the 1:1 Languir model is used for kinetic analysis.

[0025] In some embodiments of any of the foregoing aspects, the antibody can inhibit the enzymatic activity of human tryptase beta 1. In some embodiments, the antibody inhibits the activity of tryptase with an IC50 of about 2.5 nM or less as determined by a human tryptase beta enzyme assay using synthetic peptide S-2288 as a substrate. In some embodiments, the antibody inhibits the activity of tryptase with an IC50 of about 550 pM to about 2.5 nM. In some embodiments, the antibody inhibits the activity of tryptase with an IC50 of about 500 pM to about 2 nM. In some embodiments, the antibody inhibits the activity of tryptase with an IC50 of about 550 nM to 1.5 nM. In some embodiments, the antibody inhibits the activity of tryptase with an IC50 of about 500 pM to about 700 pM. In some embodiments, the inhibitory activity of the antibody is determined as described in Example 1(A)(viii)(a). In some embodiments, the final concentration of heparin in the human tryptase beta enzyme assay using synthetic peptide S-2288 is 66 μg / ml. In some embodiments, recombinant human tryptase beta 1 tetramer active enzyme is diluted to 0.75 nM in TNH buffer (200 mM Tris, 150 mM NaCl, 0.1 mg / mL heparin, 0.01% TRITON™ X-100, pH 8.0) and combined 1:1 with anti-tryptase antibody (diluted in PBS) in a 384-well plate. The plate is incubated for 1 hour at ambient temperature with gentle agitation. Colorimetric substrate S-2288 (Chromogenix, product number 82-0852-39), or an equivalent substrate, is diluted to 1200 μM in TNH buffer and added to the plate. In some embodiments, the final well concentrations are 400 μM S-2288, 0.25 nM recombinant human tryptase beta 1 tetramer, 66 μg / mL heparin, and 0.10 - ②②② nM anti-tryptase antibody. The plate is incubated for 40 minutes at ambient temperature with gentle agitation and then read at A 405 Read at. The IC50 of the anti-tryptase antibody is determined from the four-parameter fit of their respective curves.

[0026] In some embodiments of any of the foregoing aspects, the antibody can inhibit the enzymatic activity of human tryptase beta 1 at pH 6. Specifically, the inhibitory activity of the antibody can be determined at pH 6.

[0027] In some embodiments, the antibody can inhibit tryptase-mediated stimulation of bronchial smooth muscle cell proliferation and / or collagen-based contraction. In some embodiments, the antibody can inhibit mast cell histamine release. In some embodiments, the antibody can inhibit IgE-induced histamine release and / or tryptase-induced histamine release. In some embodiments, the antibody can inhibit cynomolgus tryptase D1 as evaluated by an active tryptase ELISA assay. In some embodiments, the antibody can inhibit tryptase activity in cynomolgus bronchoalveolar lavage (BAL) or nasal absorption samples. In some embodiments, the antibody can dissociate tetrameric human tryptase beta 1. In some embodiments, the antibody can dissociate tetrameric human tryptase beta 1 when in a monovalent format. In some embodiments, the monovalent format is the Fab format. In some embodiments, the antibody can dissociate tetrameric human tryptase beta 1 in the presence of heparin. In some embodiments, the antibody can dissociate tetrameric tryptase beta in the presence of 66 μg / ml of heparin.

[0028] In another aspect, the invention features an antibody that binds to the same epitope as any one of the aforementioned antibodies. In some embodiments, whether the antibody binds to the same epitope or competes for binding to human tryptase beta 1 is determined by an epitope binning assay. In some embodiments, the epitope binning assay is an OCTET® epitope binning assay as described in Section C of Example 3. In some embodiments, the human tryptase beta 1 monomeric protein is biotinylated by reacting with NHS-PEG4 biotin at Lys residues. The biotinylated monomer is diluted at 5 μg / ml with running buffer (ForteBio, Inc.) and immobilized on a streptavidin sensor chip (ForteBio, Inc.). After the immobilization step, the human tryptase beta 1-immobilized sensor is saturated with the first antibody, diluted at 10-20 μg / ml, and then bound with the second antibody diluted at 2.5 μg / ml. In some embodiments, the epitope binning assay is performed at 30°C.

[0029] In another aspect, the invention features an antibody that competes for binding to human tryptase beta 1 with any one of the aforementioned antibodies, or cross-blocks it, or is cross-blocked thereby.

[0030] In some embodiments of any of the aforementioned aspects, the antibody is monoclonal, human, humanized, or chimeric. In some embodiments, the antibody is humanized.

[0031] In some embodiments of any of the aforementioned aspects, the antibody is an antibody fragment that binds to tryptase. In some embodiments, the antibody fragment is selected from the group consisting of Fab, Fab'-SH, Fv, scFv, and (Fab')2 fragments.

[0032] In some embodiments of any of the foregoing aspects, the antibody is a full-length antibody. In some embodiments, the antibody is an IgG antibody. In some embodiments, the IgG antibody is an IgG1 antibody. In some embodiments, the IgG antibody is an IgG4 antibody. In some embodiments, the IgG4 antibody comprises a mutation in the hinge region. In some embodiments, the mutation is a substitution mutation. In some embodiments, the substitution mutation is at amino acid residue S228 (EU numbering). In some embodiments, the IgG4 antibody comprises the S228P mutation (EU numbering).

[0033] In some embodiments of any of the above aspects, the antibody is a monospecific antibody.

[0034] In some embodiments of any of the foregoing aspects, the antibody is a multispecific antibody. In some embodiments, the antibody is a bispecific antibody. In some embodiments, the antibody comprises a first binding domain that binds to tryptase and a second binding domain that binds to a second biological molecule, wherein the second biological molecule is selected from the group consisting of interleukin-13 (IL-13), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-17 (IL-17), IgE, and interleukin-33 (IL-33). In some embodiments, the second biological molecule is IL-13. In some embodiments, the second biological molecule is IL-33. In some embodiments, the second biological molecule is IgE.

[0035] In another aspect, the invention features an isolated nucleic acid encoding any of the antibodies described herein, or a set of isolated nucleic acids encoding the antibodies together.

[0036] In another aspect, the present invention features an isolated nucleic acid encoding an antibody comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 9 and / or a VL domain comprising the amino acid sequence of SEQ ID NO: 10, or a set of isolated nucleic acids encoding the antibody together, wherein the nucleic acid comprises a sequence that is at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence of SEQ ID NO: 104 and / or SEQ ID NO: 105. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76 and / or (b) a light chain comprising the amino acid sequence of SEQ ID NO: 77, and the nucleic acid or set comprises a sequence that is at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence of SEQ ID NO: 106 and / or SEQ ID NO: 107. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 78 and / or (b) a light chain comprising the amino acid sequence of SEQ ID NO: 79, and the nucleic acid or set comprises a sequence that is at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence of SEQ ID NO: 108 and / or SEQ ID NO: 107. In some embodiments, the nucleic acid or set comprises the sequence of SEQ ID NO: 108 and / or SEQ ID NO: 107.

[0037] In another aspect, the present invention features an isolated nucleic acid encoding an antibody comprising a VH domain comprising the amino acid sequence of SEQ ID NO: 36 and / or a VL domain comprising the amino acid sequence of SEQ ID NO: 37, or a set of isolated nucleic acids encoding the antibody together, wherein the nucleic acid comprises a sequence that is at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence of SEQ ID NO: 109 and / or SEQ ID NO: 110. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and / or (b) a light chain comprising the amino acid sequence of SEQ ID NO: 81, and the nucleic acid or set comprises a sequence that is at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence of SEQ ID NO: 111 and / or SEQ ID NO: 112. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and / or (b) a light chain comprising the amino acid sequence of SEQ ID NO: 83, and the nucleic acid or set comprises a sequence that is at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence of SEQ ID NO: 113 and / or SEQ ID NO: 112. In some embodiments, the nucleic acid or set comprises the sequence of SEQ ID NO: 113 and / or SEQ ID NO: 112.

[0038] In another aspect, the present invention features a vector (e.g., an expression vector) or a set of vectors comprising any of the isolated nucleic acids or sets of isolated nucleic acids described herein. In another aspect, the present invention features a host cell comprising the aforementioned nucleic acid and / or vector and / or set of nucleic acids and / or set of vectors. In some embodiments, the host cell is a mammalian cell. In some embodiments, the mammalian cell is a Chinese hamster ovary (CHO) cell. In some embodiments, the host cell is a prokaryotic cell. In some embodiments, the prokaryotic cell is E. coli.

[0039] In another aspect, the present invention is a method for producing any one of the antibodies described herein, comprising culturing a host cell containing any one of the aforementioned vectors (e.g., expression vectors) or a set of vectors in a culture medium under suitable conditions that enable the production of the antibody. In some embodiments, the method further comprises recovering the antibody from the host cell or the culture medium.

[0040] In another aspect, the present invention features a composition (e.g., a pharmaceutical composition) comprising any one of the aforementioned antibodies. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent.

[0041] In another aspect, the present invention features a pharmaceutical composition comprising an isolated monoclonal antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, and a pharmaceutically acceptable carrier, excipient, or diluent, wherein the antibody binds to monomeric tryptase beta 1 with a K of about 0.1 nM to about 1 nM and / or the antibody can inhibit the enzymatic activity of tryptase with a half-maximal inhibitory concentration (IC50) of about 0.1 nM to about 5 nM as determined by an in vitro tryptase enzyme assay using S-2288 as a substrate. D and / or the antibody can inhibit the enzymatic activity of tryptase with a half-maximal inhibitory concentration (IC50) of about 0.1 nM to about 5 nM.

[0042] In some embodiments of the aforementioned aspects, the antibody binds to tryptase with a K of about 0.5 nM to about 1 nM. D In some embodiments, the antibody binds to tryptase with a K of about 0.1 nM to about 0.5 nM. D In some embodiments, the antibody binds to tryptase with a K of about 0.4 nM. D In some embodiments, the antibody binds to tryptase with a K of about 0.2 nM. D In some embodiments, the K D is measured by a surface plasmon resonance (SPR) assay. In some embodiments, the SPR assay is performed at 25°C. In some embodiments, the K Dis measured, for example, using a BIACORE® SPR assay as described in section (A)(vii) of Example 1. In some embodiments, the SPR assay can use a BIAcore® T200 or equivalent instrument. In some embodiments, a BIAcore® Series S CM5 sensor chip (or equivalent sensor chip) is immobilized with a monoclonal mouse anti-human IgG (Fc) antibody, and the anti-tryptase antibody is sequentially captured on the flow cell. Serial three-fold dilutions of His-tagged human tryptase beta 1 monomer (SEQ ID NO: 128) are injected at a flow rate of 30 μl / min. Each sample is analyzed for 3 minutes association and 10 minutes dissociation. The assay is performed at 25° C. After each injection, the chip is regenerated using 3 M MgCl2. The binding response is corrected by subtracting the response units (RU) from a flow cell that captures an unrelated IgG of similar density. k on and k off The simultaneous fitting of a 1:1 Languir model for is used for kinetic analysis.

[0043] In some embodiments of the foregoing aspects, the antibody can inhibit the activity of tryptase with an IC50 of about 0.5 nM to about 5 nM. In some embodiments, the antibody can inhibit the activity of tryptase with an IC50 of about 0.1 nM to about 2 nM. In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 4 nM. In some embodiments, the antibody can inhibit the activity of tryptase with an IC50 of about 0.6 nM. In some embodiments, the antibody can inhibit tryptase activity at pH 6 in an in vitro tryptase enzyme assay using S-2288 as a substrate. In some embodiments, the inhibitory activity of the antibody is determined as described in the Examples (e.g., Example 1, Section (A)(viii)(a)). In some embodiments, recombinant human tryptase beta1 tetramer active enzyme is diluted to 0.75 nM in TNH buffer (200 mM Tris, 150 mM NaCl, 0.1 mg / mL heparin, 0.01% TRITON™ X-100, pH 8.0) and combined 1:1 with anti-tryptase antibody (diluted in PBS) in a 384-well plate. The plate is incubated for 1 hour at ambient temperature with gentle agitation. The colorimetric substrate S-2288 (Chromogenix, product number 82-0852-39), or an equivalent substrate, is diluted to 1200 μM in TNH buffer and added to the plate. In some embodiments, the final well concentrations are 400 μM S-2288, 0.25 nM recombinant human tryptase beta1 tetramer, 66 μg / mL heparin, and 0.10 - 222 nM anti-tryptase antibody. The plate is incubated for 40 minutes at ambient temperature with gentle agitation and then read at A 405 The IC50 of the anti-tryptase antibodies is determined from a four-parameter fit of their respective curves. In some embodiments, the final concentration of heparin in the human tryptase beta enzyme assay using the synthetic peptide S-2288 is 66 μg / ml.

[0044] In some embodiments of the foregoing aspects, the antibody can inhibit tryptase-mediated stimulation of bronchial smooth muscle cell proliferation and / or collagen-based contraction. In some embodiments, the antibody can inhibit mast cell histamine release. In some embodiments, the antibody can inhibit IgE-induced histamine release and / or tryptase-induced histamine release. In some embodiments, the antibody can inhibit tryptase activity in cynomolgus bronchial alveolar lavage (BAL) or nasal absorption samples. In some embodiments, the antibody can dissociate tetrameric human tryptase beta 1. In some embodiments, the antibody can dissociate tetrameric human tryptase beta 1 when in a monovalent format. In some embodiments, the monovalent format is the Fab format. In some embodiments, the antibody can dissociate tetrameric human tryptase beta 1 in the presence of heparin. In some embodiments, the antibody can dissociate tetrameric tryptase beta in the presence of 66 μg / ml of heparin.

[0045] In some embodiments of the foregoing aspects, the antibody comprises the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8), (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some embodiments, the antibody comprises (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 9, (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% identity with the amino acid sequence of SEQ ID NO: 10, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFS (SEQ ID NO: 11), (b) FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 12), (c) FR-H3 comprising the amino acid sequence of RFTISRDNSKNTLYLQMNSLRAEDTAVYYCTR (SEQ ID NO: 13), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14). In some embodiments, the VH domain of the antibody comprises the amino acid sequence of SEQ ID NO: 9. In some embodiments, the antibody further comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 15), (b) FR-L2 comprising the amino acid sequence of WYQQKPGKSPKPWIY (SEQ ID NO: 16), (c) FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 17), and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 18).In some embodiments, the VL domain of the antibody comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 77. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 78 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 79. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 comprising at least 1, at least 2, at least 3, or all 4 residues selected from the group consisting of His51, Val80, Lys81, and Asp82 of SEQ ID NO: 71. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 comprising His51 of SEQ ID NO: 71 and at least 1, at least 2, or all 3 residues selected from the group consisting of Val80, Lys81, and Asp82. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or all 12 amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises His51, Gln67, Val80, Lys81, Asp82, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope is related to the human tryptase beta 1 monomer or tetramer.In some embodiments, the epitope is determined by an X-ray crystallographic model. In some embodiments, the antibody can dissociate both the small interface of tetrameric human tryptase beta 1 and the large interface of tetrameric human tryptase beta 1.

[0046] In other embodiments of the foregoing aspects, the antibody comprises the following six HVRS: (a) HVR-H1 comprising the amino acid sequence of GYAIT (SEQ ID NO: 30), (b) HVR-H2 comprising the amino acid sequence of GISSAATTFYSSWAKS (SEQ ID NO: 31), (c) HVR-H3 comprising the amino acid sequence of DPRGYGAALDRLDL (SEQ ID NO: 32), (d) HVR-L1 comprising the amino acid sequence of QSIKSVYNNRLG (SEQ ID NO: 33), (e) HVR-L2 comprising the amino acid sequence of ETSILTS (SEQ ID NO: 34), and (f) HVR-L3 comprising the amino acid sequence of AGGFDRSGDTT (SEQ ID NO: 35). In some embodiments, the antibody comprises (a) a VH domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 36, 47, 48, 49, 50, 51, and 52, (b) a VL domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% identity with any one of the amino acid sequences of SEQ ID NOs: 37, 53, 58, or 59, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVQLVESGPGLVKPSETLSLTCTVSRFSLI (SEQ ID NO: 38), (b) FR-H2 comprising the amino acid sequence of WIRQPPGKGLEWIG (SEQ ID NO: 42), (c) FR-H3 comprising the amino acid sequence of RVTISRDTSKNQVSLKLSSVTAADTAVYYCAR (SEQ ID NO: 43), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41). In some embodiments, the VH domain of the antibody comprises the amino acid sequence of SEQ ID NO: 36.In some embodiments, the antibody further comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 64), (b) FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 65), (c) FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 66), and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 63). In some embodiments, the VL domain of the antibody comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 81. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 83. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 comprising at least 1, at least 2, or all 3 residues selected from the group consisting of Gln100, Leu101, and Leu102 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or all 14 amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71.In some embodiments, the epitope comprises Gln35, Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Gln100, Leu101, Leu102, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, Glu129, and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is related to the human tryptase beta 1 monomer or tetramer. In some embodiments, the epitope is related to the human tryptase beta 1 tetramer, and the epitope on human tryptase beta 1 further comprises one or both of Gln35 and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is determined by an X-ray crystallography model. In some embodiments, the antibody can dissociate the small interface and / or the large interface of human tryptase beta 1.

[0047] In another aspect, the invention features a composition (e.g., a pharmaceutical composition) comprising an isolated monoclonal antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, and a pharmaceutically acceptable carrier, excipient, or diluent, wherein the antibody binds to an epitope on human tryptase beta 1 that comprises at least 1, at least 2, at least 3, or at least 4 all of the residues selected from the group consisting of His51, Val80, Lys81, and Asp82 of SEQ ID NO: 71. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 that comprises His51 of SEQ ID NO: 71 and at least 1, at least 2, or all 3 of the residues selected from the group consisting of Val80, Lys81, and Asp82. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or all 12 of the amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises His51, Gln67, Val80, Lys81, Asp82, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope is associated with the human tryptase beta 1 monomer or tetramer. In some embodiments, the epitope is determined by an X-ray crystallography model.In some embodiments, the antibody can dissociate both the small interface of tetrameric human tryptase beta1 and the large interface of tetrameric human tryptase beta1. In some embodiments, the antibody comprises the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8), (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some embodiments, the antibody comprises (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity with the amino acid sequence of SEQ ID NO: 9, (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% identity with the amino acid sequence of SEQ ID NO: 10, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFS (SEQ ID NO: 11), (b) FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 12), (c) FR-H3 comprising the amino acid sequence of RFTISRDNSKNTLYLQMNSLRAEDTAVYYCTR (SEQ ID NO: 13), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14). In some embodiments, the VH domain of the antibody comprises the amino acid sequence of SEQ ID NO: 9.In some embodiments, the antibody further comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 15), (b) FR-L2 comprising the amino acid sequence of WYQQKPGKSPKPWIY (SEQ ID NO: 16), (c) FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 17), and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 18). In some embodiments, the VL domain of the antibody comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 77. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 78 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 79.

[0048] In another aspect, the present invention features a composition (e.g., a pharmaceutical composition) comprising an isolated monoclonal antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, and a pharmaceutically acceptable carrier, excipient, or diluent, wherein the antibody binds to an epitope on human tryptase beta 1 that comprises at least 1, at least 2, or all 3 residues selected from the group consisting of Gln100, Leu101, and Leu102 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or all 14 amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71. In some embodiments, the epitope comprises Gln35, Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Gln100, Leu101, Leu102, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, Glu129, and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is associated with the human tryptase beta 1 monomer or tetramer. In some embodiments, the epitope is associated with the human tryptase beta 1 tetramer and the epitope on human tryptase beta 1 further comprises one or both of Gln35 and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is determined by an X-ray crystallographic model.In some embodiments, the antibody can dissociate the small and / or large interfaces of human tryptase beta 1. In some embodiments, the antibody comprises the following six HVRS: (a) HVR-H1 comprising the amino acid sequence of GYAIT (SEQ ID NO: 30), (b) HVR-H2 comprising the amino acid sequence of GISSAATTFYSSWAKS (SEQ ID NO: 31), (c) HVR-H3 comprising the amino acid sequence of DPRGYGAALDRLDL (SEQ ID NO: 32), (d) HVR-L1 comprising the amino acid sequence of QSIKSVYNNRLG (SEQ ID NO: 33), (e) HVR-L2 comprising the amino acid sequence of ETSILTS (SEQ ID NO: 34), and (f) HVR-L3 comprising the amino acid sequence of AGGFDRSGDTT (SEQ ID NO: 35). In some embodiments, the antibody comprises (a) a VH domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% sequence identity with any one of the amino acid sequences of SEQ ID NOs: 36, 47, 48, 49, 50, 51, and 52, (b) a VL domain comprising an amino acid sequence having at least 90%, at least 95%, or at least 99% identity with any one of the amino acid sequences of SEQ ID NOs: 37, 53, 58, or 59, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody further comprises the following VH domain FRs: (a) FR-H1 comprising the amino acid sequence of EVQLVESGPGLVKPSETLSLTCTVSRFSLI (SEQ ID NO: 38), (b) FR-H2 comprising the amino acid sequence of WIRQPPGKGLEWIG (SEQ ID NO: 42), (c) FR-H3 comprising the amino acid sequence of RVTISRDTSKNQVSLKLSSVTAADTAVYYCAR (SEQ ID NO: 43), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41). In some embodiments, the VH domain of the antibody comprises the amino acid sequence of SEQ ID NO: 36.In some embodiments, the antibody further comprises the following VL domain FRs: (a) FR-L1 comprising the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 64), (b) FR-L2 comprising the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 65), (c) FR-L3 comprising the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 66), and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 63). In some embodiments, the VL domain of the antibody comprises the amino acid sequence of SEQ ID NO: 37. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 81. In some embodiments, the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 83.

[0049] In some embodiments, the antibody can further bind to human tryptase alpha, tryptase beta 2, tryptase beta 3, and / or cynomolgus tryptase D1.

[0050] In any of the foregoing compositions (e.g., pharmaceutical compositions), the antibody can be monoclonal, human, humanized, or chimeric. In some embodiments, the antibody is humanized.

[0051] In any of the foregoing compositions (e.g., pharmaceutical compositions), the composition can be for use in humans.

[0052] Any of the foregoing compositions (e.g., pharmaceutical compositions) can be lyophilized. In other embodiments, the foregoing compositions (e.g., pharmaceutical compositions) can be liquid.

[0053] The excipient of the aforementioned composition (e.g., pharmaceutical composition) can be an antioxidant. In some embodiments, the composition comprises one or more antioxidants selected from the group consisting of N-acetyltryptophan, tryptophan, methionine, cysteine, glutathione, thiosorbitol, ascorbic acid, monothioglycerol, cyclodextrin, trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), pyridoxine, mannitol, and metal chelating agents. In some embodiments, the composition comprises N-acetyltryptophan or methionine. In some embodiments, the composition comprises N-acetyltryptophan and methionine.

[0054] In another aspect, the present invention features a composition (e.g., a pharmaceutical composition) comprising (i) an isolated antibody that binds to human tryptase beta 1, or an antigen-binding fragment thereof, the antibody comprising the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8), (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6), wherein the oxidation of tryptophan at position 6 of HVR-H3 (SEQ ID NO: 8) is 30% or less. In some embodiments, the oxidation of tryptophan at position 6 of HVR-H3 (SEQ ID NO: 8) is 28% or less, 25% or less, 20% or less, 15% or less, 10% or less, or 6% or less. In some embodiments, the oxidation of tryptophan at position 6 of HVR-H3 (SEQ ID NO: 8) is determined according to the AAPH stress test. In some embodiments, the oxidation of tryptophan at position 6 of HVR-H3 (SEQ ID NO: 8) is determined within one year from the initial production of the composition.

[0055] Any of the foregoing compositions (e.g., pharmaceutical compositions) may contain N-acetyltryptophan at a concentration of about 0.1 mM to about 5 mM. In some embodiments, the concentration of N-acetyltryptophan is about 0.1 mM to about 1 mM. In some embodiments, the concentration of N-acetyltryptophan is about 0.3 mM. In some embodiments, the composition contains methionine at a concentration of about 1 mM to about 20 mM. In some embodiments, the concentration of methionine is about 1 mM to about 10 mM. In some embodiments, the concentration of methionine is about 5 mM.

[0056] In another aspect, the present invention features a composition (e.g., a pharmaceutical composition) comprising: (i) an isolated antibody or antigen-binding fragment thereof that binds to human tryptase, wherein the antibody comprises the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8), (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6); (ii) N-acetyltryptophan at a concentration of about 0.1 mM to about 1 mM; and (iii) methionine at a concentration of about 1 mM to about 10 mM.

[0057] In any of the foregoing compositions (e.g., pharmaceutical compositions), the concentration of the antibody can be about 1 mg / ml to about 250 mg / ml. In some embodiments, the concentration of the antibody is about 150 mg / ml.

[0058] Any of the foregoing compositions (e.g., pharmaceutical compositions) may further comprise one or more additional excipients selected from the group consisting of stabilizers, buffers, surfactants, and tonicity agents. In some embodiments, the composition further comprises a buffer. In some embodiments, the buffer is arginine succinate and / or histidine succinate. In some embodiments, the buffer comprises arginine succinate and histidine succinate. In some embodiments, the concentration of arginine succinate is from about 50 mM to about 500 mM. In some embodiments, the concentration of arginine succinate is from about 100 mM to about 300 mM. In some embodiments, the concentration of arginine succinate is about 200 mM. In some embodiments, the concentration of histidine succinate is from about 1 mM to about 50 mM. In some embodiments, the concentration of histidine succinate is from about 15 mM to about 25 mM. In some embodiments, the concentration of histidine succinate is about 20 mM. In some embodiments, the composition further comprises a surfactant. In some embodiments, the surfactant is poloxamer 188 or polysorbate 20. In some embodiments, the surfactant is poloxamer 188. In some embodiments, the concentration of poloxamer 188 is from about 0.005% to about 0.1%. In some embodiments, the concentration of poloxamer 188 is from about 0.005% to about 0.05%. In some embodiments, the concentration of poloxamer 188 is about 0.02%. In some embodiments, the pH of the composition is from about 4.5 to about 7.0. In some embodiments, the pH of the composition is from about 4.5 to about 6.5. In some embodiments, the pH of the composition is about 5.5. In some embodiments, the composition is in a light-protective container. In some embodiments, the composition is in a prefilled syringe.

[0059] Any of the foregoing compositions (e.g., pharmaceutical compositions) may further comprise an IL-13 axis-binding antagonist, an IL-5 axis-binding antagonist, an IL-33 axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor. In some embodiments, the IL-13 axis-binding antagonist is an anti-IL-13 antibody. In some embodiments, the anti-IL-13 antibody is lebrikizumab. In some embodiments, the IL-5 axis-binding antagonist is an IL-5 binding antagonist or an IL-5 receptor binding antagonist. In some embodiments, the IL-33 axis-binding antagonist is an IL-33 binding antagonist or an ST2 binding antagonist. In some embodiments, the IL-33 binding antagonist is an anti-IL-33 antibody. In some embodiments, the M1 prime antagonist is kirizumab. In some examples, the IgE antagonist is omalizumab (XOLAIR®).

[0060] Any of the foregoing compositions (e.g., pharmaceutical compositions) may be formulated for administration to a human. In certain embodiments, the pharmaceutical composition comprises an antibody that does not contain a non-human constant region sequence. In certain embodiments, the pharmaceutical composition comprises an antibody that does not contain a non-human framework and a non-human constant region sequence. In certain embodiments, the pharmaceutical composition comprises an antibody that is a human antibody, a humanized antibody, or a chimeric antibody.

[0061] In some aspects, any one of the foregoing antibodies may be used as a medicament.

[0062] In some embodiments, any one of the aforementioned antibodies can be used in treating a disorder. In some embodiments, the disorder is selected from the group consisting of a pulmonary disorder, an immunodeficiency, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, a lymphocytic disorder, or a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is a pulmonary disorder. In some embodiments, the pulmonary disorder is selected from the group consisting of asthma, airway hyperreactivity, and chronic obstructive pulmonary disease (COPD). In some embodiments, the pulmonary disorder is asthma. In some embodiments, the asthma is Th2-high asthma or Th2-low asthma. In some embodiments, the immunodeficiency is selected from the group consisting of rheumatoid arthritis, psoriasis, eosinophilic esophagitis, inflammatory bowel disease (IBD), and Crohn's disease. In some embodiments, the inflammatory disorder is chronic idiopathic urticaria (CIU, also known as chronic spontaneous urticaria, CSU), anaphylaxis, anaphylactic shock, atopic dermatitis, or allergic rhinitis. In some embodiments, the fibrotic disorder is idiopathic pulmonary fibrosis (IPF). In some embodiments, the disorder is a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is mastocytosis. In some embodiments, the antibody is for use in combination with a further therapeutic agent. In some embodiments, the further therapeutic agent is an IL-13 axis-binding antagonist, an IL-5 axis-binding antagonist, an IL-33 axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor.In some embodiments, the IL-13 axis-binding antagonist is an anti-IL-13 antibody. In some embodiments, the anti-IL-13 antibody is lebrikizumab. In some embodiments, the IL-5 axis-binding antagonist is an IL-5 binding antagonist or an IL-5 receptor binding antagonist. In some embodiments, the IL-33 axis-binding antagonist is an IL-33 binding antagonist or an ST2 binding antagonist. In some embodiments, the IL-33 binding antagonist is an anti-IL-33 antibody. In some embodiments, the M1 prime antagonist is kirizumab. In some embodiments, the antibody is for administration subcutaneously, intravenously, intramuscularly, topically, orally, transdermally, intraperitoneally, intraorbitally, implanted, inhaled, intrathecally, intravesically, or intranasally. In some embodiments, the antibody is for subcutaneous administration. In some embodiments, the antibody is for use in a human subject.

[0063] In some aspects, any one of the foregoing compositions (e.g., pharmaceutical compositions) can be used as a medicament.

[0064] In some embodiments, any of the foregoing compositions (e.g., pharmaceutical compositions) can be used in treating a disorder. In some embodiments, the disorder is selected from the group consisting of a lung disorder, an autoimmune disorder, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, a lymphocytic disorder, or a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is a lung disorder. In some embodiments, the lung disorder is selected from the group consisting of asthma, airway hyperreactivity, and chronic obstructive pulmonary disease (COPD). In some embodiments, the lung disorder is asthma. In some embodiments, the asthma is Th2-high asthma or Th2-low asthma. In some embodiments, the autoimmune disorder is selected from the group consisting of rheumatoid arthritis, psoriasis, eosinophilic esophagitis, inflammatory bowel disease (IBD), and Crohn's disease. In some embodiments, the inflammatory disorder is chronic idiopathic urticaria (CIU or CSU), anaphylaxis, anaphylactic shock, atopic dermatitis, or allergic rhinitis. In some embodiments, the fibrotic disorder is idiopathic pulmonary fibrosis (IPF). In some embodiments, the disorder is a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is mastocytosis. In some embodiments, the composition (e.g., pharmaceutical composition) is for use in combination with a further therapeutic agent. In some embodiments, the further therapeutic agent is an IL-13 axis-binding antagonist, an IL-5 axis-binding antagonist, an IL-33 axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor.In some embodiments, the IL-13 axis-binding antagonist is an anti-IL-13 antibody. In some embodiments, the anti-IL-13 antibody is lebrikizumab. In some embodiments, the IL-5 axis-binding antagonist is an IL-5 binding antagonist or an IL-5 receptor binding antagonist. In some embodiments, the IL-33 axis-binding antagonist is an IL-33 binding antagonist or an ST2 binding antagonist. In some embodiments, the IL-33 binding antagonist is an anti-IL-33 antibody. In some embodiments, the M1 prime antagonist is kirizumab. In some embodiments, the composition (e.g., pharmaceutical composition) is for administration subcutaneously, intravenously, intramuscularly, topically, orally, transdermally, intraperitoneally, intravitreally, implantably, by inhalation, intrathecally, intravesically, or intranasally. In some embodiments, the composition (e.g., pharmaceutical composition) is administered subcutaneously. In some embodiments, the composition (e.g., pharmaceutical composition) is for use in a human subject.

[0065] In some embodiments, any one of the foregoing antibodies can be used in the manufacture of a medicament for treating a disorder. In some embodiments, the disorder is selected from the group consisting of a pulmonary disorder, an autoimmune disorder, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, a lymphocytic disorder, or a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is a pulmonary disorder. In some embodiments, the pulmonary disorder is selected from the group consisting of asthma, airway hyperreactivity, and chronic obstructive pulmonary disease (COPD). In some embodiments, the pulmonary disorder is asthma. In some embodiments, the asthma is Th2-high asthma or Th2-low asthma. In some embodiments, the autoimmune disorder is selected from the group consisting of rheumatoid arthritis, psoriasis, eosinophilic esophagitis, inflammatory bowel disease (IBD), and Crohn's disease. In some embodiments, the inflammatory disorder is chronic idiopathic urticaria (CIU or CSU), anaphylaxis, anaphylactic shock, atopic dermatitis, or allergic rhinitis. In some embodiments, the fibrotic disorder is idiopathic pulmonary fibrosis (IPF). In some embodiments, the disorder is a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is mastocytosis. In some embodiments, the medicament is formulated for use in combination with a further therapeutic agent. In some embodiments, the further therapeutic agent is an IL-13 axis-binding antagonist, an IL-5 axis-binding antagonist, an IL-33 axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor. In some embodiments, the IL-13 axis-binding antagonist is an anti-IL-13 antibody.In some embodiments, the anti-IL-13 antibody is lebrikizumab. In some embodiments, the IL-5 axis-binding antagonist is an IL-5 binding antagonist or an IL-5 receptor binding antagonist. In some embodiments, the IL-33 axis-binding antagonist is an IL-33 binding antagonist or an ST2 binding antagonist. In some embodiments, the IL-33 binding antagonist is an anti-IL-33 antibody. In some embodiments, the M1 prime antagonist is kirizumab. In some embodiments, the pharmaceutical is formulated for administration subcutaneously, intravenously, intramuscularly, topically, orally, transdermally, intraperitoneally, intravitreally, implanted, by inhalation, intrathecally, intravesically, or intranasally. In some embodiments, the pharmaceutical is formulated for subcutaneous administration. In some embodiments, the pharmaceutical is formulated for use in a human subject.

[0066] In some embodiments, any one of the foregoing compositions (e.g., pharmaceutical compositions) can be used in the manufacture of a medicament for treating a disorder. In some embodiments, the disorder is selected from the group consisting of a lung disorder, an autoimmune disorder, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, a lymphocytic disorder, or a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is a lung disorder. In some embodiments, the lung disorder is selected from the group consisting of asthma, airway hyperreactivity, and chronic obstructive pulmonary disease (COPD). In some embodiments, the lung disorder is asthma. In some embodiments, the asthma is Th2-high asthma or Th2-low asthma. In some embodiments, the autoimmune disorder is selected from the group consisting of rheumatoid arthritis, psoriasis, eosinophilic esophagitis, inflammatory bowel disease (IBD), and Crohn's disease. In some embodiments, the inflammatory disorder is chronic idiopathic urticaria (CIU or CSU), anaphylaxis, anaphylactic shock, atopic dermatitis, or allergic rhinitis. In some embodiments, the fibrotic disorder is idiopathic pulmonary fibrosis (IPF). In some embodiments, the disorder is a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is mastocytosis. In some embodiments, the medicament is formulated for use in combination with a further therapeutic agent. In some embodiments, the further therapeutic agent is an IL-13 axis-binding antagonist, an IL-5 axis-binding antagonist, an IL-33 axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor.In some embodiments, the IL-13 axis-binding antagonist is an anti-IL-13 antibody. In some embodiments, the anti-IL-13 antibody is lebrikizumab. In some embodiments, the IL-5 axis-binding antagonist is an IL-5 binding antagonist or an IL-5 receptor binding antagonist. In some embodiments, the IL-33 axis-binding antagonist is an IL-33 binding antagonist or an ST2 binding antagonist. In some embodiments, the IL-33 binding antagonist is an anti-IL-33 antibody. In some embodiments, the M1 prime antagonist is kirizumab. In some embodiments, the pharmaceutical is formulated for administration subcutaneously, intravenously, intramuscularly, topically, orally, transdermally, intraperitoneally, intraorbitally, implanted, inhaled, intrathecally, intravesically, or intranasally. In some embodiments, the pharmaceutical is formulated for subcutaneous administration. In some embodiments, the pharmaceutical is formulated for use in a human subject.

[0067] In another aspect, the present invention features a method of treating a disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the aforementioned antibodies. In some embodiments, the disorder is selected from the group consisting of a pulmonary disorder, an immunodeficiency, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, a lymphocytic disorder, or a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is a pulmonary disorder. In some embodiments, the pulmonary disorder is selected from the group consisting of asthma, airway hyperreactivity, and chronic obstructive pulmonary disease (COPD). In some embodiments, the pulmonary disorder is asthma. In some embodiments, the asthma is Th2-high asthma or Th2-low asthma. In some embodiments, the immunodeficiency is selected from the group consisting of rheumatoid arthritis, psoriasis, eosinophilic esophagitis, inflammatory bowel disease (IBD), and Crohn's disease. In some embodiments, the inflammatory disorder is chronic idiopathic urticaria (CIU or CSU), anaphylaxis, anaphylactic shock, atopic dermatitis, or allergic rhinitis. In some embodiments, the fibrotic disorder is idiopathic pulmonary fibrosis (IPF). In some embodiments, the disorder is a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is mastocytosis. In some embodiments, the method further comprises administering a further therapeutic agent to the subject. In some embodiments, the further therapeutic agent is an IL-13 axis-binding antagonist, an IL-5 axis-binding antagonist, an IL-33 axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor.In some embodiments, the IL-13 axis-binding antagonist is an anti-IL-13 antibody. In some embodiments, the anti-IL-13 antibody is lebrikizumab. In some embodiments, the IL-5 axis-binding antagonist is an IL-5 binding antagonist or an IL-5 receptor binding antagonist. In some embodiments, the IL-33 axis-binding antagonist is an IL-33 binding antagonist or an ST2 binding antagonist. In some embodiments, the IL-33 binding antagonist is an anti-IL-33 antibody. In some embodiments, the M1 prime antagonist is kirizumab. In some embodiments, the IgE antagonist is omalizumab (Xolair®). In some embodiments, the antibody is administered subcutaneously, intravenously, intramuscularly, topically, orally, transdermally, intraperitoneally, intraorbitally, implantably, by inhalation, intrathecally, intravesically, or intranasally. In some embodiments, the subject is human.

[0068] In another aspect, the present invention features a method of treating a disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the foregoing compositions (e.g., a pharmaceutical composition). In some embodiments, the disorder is selected from the group consisting of a lung disorder, an autoimmune disorder, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, a lymphocytic disorder, or a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is a lung disorder. In some embodiments, the lung disorder is selected from the group consisting of asthma, airway hyperreactivity, and chronic obstructive pulmonary disease (COPD). In some embodiments, the lung disorder is asthma. In some embodiments, the asthma is Th2-high asthma or Th2-low asthma. In some embodiments, the autoimmune disorder is selected from the group consisting of rheumatoid arthritis, psoriasis, eosinophilic esophagitis, inflammatory bowel disease (IBD), and Crohn's disease. In some embodiments, the inflammatory disorder is chronic idiopathic urticaria (CIU or CSU), anaphylaxis, anaphylactic shock, atopic dermatitis, or allergic rhinitis. In some embodiments, the fibrotic disorder is idiopathic pulmonary fibrosis (IPF). In some embodiments, the disorder is a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is mastocytosis. In some embodiments, the method further comprises administering to the subject an additional therapeutic agent. In some embodiments, the additional therapeutic agent is an IL-13 axis-binding antagonist, an IL-5 axis-binding antagonist, an IL-33 axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor.In some embodiments, the IL-13 axis-binding antagonist is an anti-IL-13 antibody. In some embodiments, the anti-IL-13 antibody is lebrikizumab. In some embodiments, the IL-5 axis-binding antagonist is an IL-5 binding antagonist or an IL-5 receptor binding antagonist. In some embodiments, the IL-33 axis-binding antagonist is an IL-33 binding antagonist or an ST2 binding antagonist. In some embodiments, the IL-33 binding antagonist is an anti-IL-33 antibody. In some embodiments, the M1 prime antagonist is omalizumab (Xolair®). In some embodiments, the composition is administered subcutaneously, intravenously, intramuscularly, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intravesically, or intranasally. In some embodiments, the subject is human.

Brief Description of the Drawings

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BRIEF DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0070] I. Definitions As used herein, the term “about” refers to the normal error range of each value readily known to those skilled in the art. References to “about” values or parameters herein include (and describe) embodiments directed to the value or parameter itself.

[0071] For the purposes of this specification, an “acceptor human framework” is a framework that includes the amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, as defined below. An “acceptor human framework” “derived from” a human immunoglobulin framework or a human consensus framework may contain the same amino acid sequence or may contain amino acid sequence variations. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the VL acceptor human framework is identical in sequence to a VL human immunoglobulin framework sequence or a human consensus framework sequence.

[0072] "Affinity" refers to the total strength of non-covalent interaction between a molecule (e.g., an antibody) and its binding partner (e.g., an antigen) at a single binding site. Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects the 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y is generally represented by the dissociation constant (K D ). Affinity can be measured by common methods known in the art including the methods described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0073] "K D is measured by surface plasmon resonance assay" when used in the context of a claim means that K D is measured according to the method described in Example 1(A)(vii), which measures the kinetic parameters of the binding of an anti-tryptase antibody to the human tryptase beta1 monomer, e.g., the His6-tagged tryptase monomer shown in SEQ ID NO: 128, and does not naturally form a tryptase tetramer. The assay can use a BIAcore® T200 or equivalent instrument. Briefly, a BIAcore® Series S CM5 sensor chip (or equivalent sensor chip) is immobilized with a monoclonal mouse anti-human IgG (Fc) antibody, and the anti-tryptase antibody is sequentially captured on the flow cell. Serial 3-fold dilutions of the human tryptase beta1 monomer are injected at a flow rate of 30 μl / min. Each sample is analyzed for 3 minutes of association and 10 minutes of dissociation. The assay is performed at 25°C. After each injection, the chip is regenerated using 3 M MgCl2. The binding response is corrected by subtracting the response units (RU) from a flow cell that captures irrelevant IgG of similar density. A 1:1 Langmuir model of simultaneous fitting of k on and k off is used for kinetic analysis.

[0074] "Affinity matured" antibodies have one or more modifications in one or more HVRs and / or framework regions that result in an improvement in the affinity of the antibody for the antigen as compared to the parental antibody that does not have those modification(s). Preferred affinity matured antibodies will have nanomolar or even picomolar affinity for the target antigen. Affinity matured antibodies are produced by procedures known in the art. For example, Marks et al. Bio / Technology 10:779-783, 1992 describes affinity maturation by VH and VL domain shuffling. Random mutagenesis of HVRs and / or framework residues is described by Barbas et al. Proc. Natl. Acad. Sci. USA 91:3809-3813, 1994, Schier et al. Gene 169:147-155, 1995, Yelton et al. J. Immunol. 155:1994-2004, 1995, Jackson et al. J. Immunol. 154(7):3310-3319, 1995, and Hawkins et al. J. Mol. Biol. 226:889-896, 1992.

[0075] As used herein, the term "antibody" is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity.

[0076] As used herein, the term "tryptase" refers to any native tryptase from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. Tryptase is also known in the art as mast cell tryptase, mast cell protease II, skin tryptase, lung tryptase, pituitary tryptase, mast cell neutral protease, and mast cell serine protease II. The term "tryptase" includes tryptase alpha (encoded in humans by TPSAB1), tryptase beta (encoded in humans by TPSAB1 and TPSB2; see below), tryptase delta (encoded in humans by TPSD1), tryptase gamma (encoded in humans by TPSG1), and tryptase epsilon (encoded in humans by PRSS22). Tryptase alpha, beta, and gamma proteins are soluble, while tryptase epsilon protein is membrane-bound. Tryptase beta and gamma are active serine proteases, but they have different specificities. Tryptase alpha and delta proteins are mostly inactive proteases because they have residues at positions that are different from those of typical active serine proteases. An exemplary tryptase alpha full-length protein sequence can be found under NCBI GenBank accession number ACZ98910.1 (SEQ ID NO: 118). An exemplary tryptase gamma full-length protein sequence can be found under Uniprot accession number Q9NRR2 or GenBank accession numbers Q9NRR2.3, AAF03695.1, NP_036599.3, or AAF76457.1. An exemplary tryptase delta full-length protein sequence can be found under Uniprot accession number Q9BZJ3 or GenBank accession number NP_036349.1. Some tryptase genes are clustered on human chromosome 16p13.3. The term includes "full-length" unprocessed tryptase, as well as any form of tryptase resulting from intracellular processing.Tryptase beta is the major tryptase expressed in mast cells, while tryptase alpha is the major tryptase expressed in basophils. Tryptase alpha and tryptase beta typically contain a leader sequence of about 30 amino acids and a catalytic sequence of about 245 amino acids (see, e.g., Schwartz, Immunol. Allergy Clin. N. Am. 26:451-463, 2006).

[0077] As used herein, the term "tryptase beta" refers to any native tryptase beta from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. Tryptase beta is a serine protease that is a major component of mast cell secretory granules. As used herein, the term encompasses tryptase beta 1 (encoded by the TPSAB1 gene, which also encodes tryptase alpha 1), tryptase beta 2 (encoded by the TPSB2 gene), and tryptase beta 3 (also encoded by the TPSB2 gene). An exemplary human tryptase beta 1 sequence is shown in SEQ ID NO:71 (see also GenBank accession number NP_003285.2). An exemplary human tryptase beta 2 sequence is shown in SEQ ID NO:72 (see also GenBank accession number AAD13876.1). An exemplary human tryptase beta 3 sequence is shown in SEQ ID NO:73 (see also GenBank accession number NP_077078.5). The term tryptase beta encompasses "full-length," unprocessed tryptase beta, as well as tryptase beta resulting from post-translational modifications, including proteolytic processing. Full-length, prototryptase beta is thought to be processed in two proteolytic steps. First, R -3Autocatalytic intermolecular cleavage occurs especially in the presence of acidic pH and polyanions (e.g., heparin or dextran sulfate). Next, the remaining propeptide is removed (most likely by dipeptidyl peptidase I). For full-length human tryptase beta 1, with respect to SEQ ID NO: 71 below, the underlined amino acid residues correspond to the native leader sequence, and the amino acid residues colored in bold and gray correspond to the prodomain, which is cleaved to form the mature protein (see, for example, Sakai et al. J. Clin. Invest. 97:988-995, 1996). TIFF0007716858000001.tif35170

[0078] Mature, enzymatically active tryptase beta is typically a homotetramer or heterotetramer, although active monomers have been reported (see, for example, Fukuoka et al. J. Immunol. 176:3165, 2006). The subunits of the tryptase beta tetramer come together by hydrophobic and polar interactions between the subunits and are stabilized by polyanions (especially heparin and dextran sulfate). The term tryptase can refer to the tryptase tetramer or the tryptase monomer. Exemplary sequences of mature human tryptase beta 1, beta 2, and beta 3 are shown in SEQ ID NO: 97, SEQ ID NO: 116, and SEQ ID NO: 117, respectively. Each subunit active site faces the central pore of the tetramer, which measures approximately 50 × 30 angstroms (see, for example, Pereira et al. Nature 392:306-311, 1998). The size of the central pore typically limits access to the active site by inhibitors. Exemplary substrates of tryptase beta include, but are not limited to, PAR2, C3, fibrinogen, fibronectin, and kininogen.

[0079] The terms "antitryptase antibody," "antibody that binds tryptase," and "antibody that specifically binds tryptase" refer to an antibody that can bind tryptase with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent when targeting tryptase. In one embodiment, the extent of binding of the antitryptase antibody to unrelated, non-tryptase proteins is less than about 10% of the binding of the antibody to tryptase, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that binds tryptase has an affinity of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., ≦10 -8 M or less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 Dissociation constant (K D In certain embodiments, the anti-tryptase antibody binds to an epitope of tryptase that is conserved among tryptases from different species.

[0080] An "antibody that binds to the same epitope" as a reference antibody refers to an antibody that contacts an overlapping set of amino acid residues of an antigen compared to the reference antibody, or that blocks the binding of the reference antibody to that antigen by 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more in a competition assay. In some embodiments, the set of amino acid residues contacted by an antibody may completely overlap or partially overlap with the set of amino acid residues contacted by the reference antibody. In some embodiments, an antibody that binds to the same epitope as a reference antibody blocks the binding of the reference antibody to that antigen by 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more in a competition assay; conversely, the reference antibody blocks the binding of the antibody to that antigen by 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more in a competition assay. Exemplary competition assays are provided herein.

[0081] In the context of the claims, the term "as determined by an epitope binding assay" means that an antibody is determined to bind to the same epitope using an OCTET® epitope binding assay as described in Section C of Example 3 and / or to compete for binding with a reference anti-tryptase antibody (e.g., hu31A.v11 or huE104.v2). Briefly, human tryptase beta 1 monomeric protein is biotinylated by reacting with NHS-PEG4 biotin at Lys residues. The biotinylated monomer is diluted to 5 μg / ml with kinetic buffer (ForteBio, Inc.) and immobilized onto a streptavidin sensor chip (ForteBio, Inc.). After the immobilization step, the human tryptase beta 1-immobilized sensor is saturated with a first antibody and bound with a second antibody diluted at 10-20 μg / ml and subsequently at 2.5 μg / ml. The test temperature for such an epitope binding assay is 30° C. A binding signal by the second antibody means that the two antibodies can bind to the antigen simultaneously at different, non-overlapping epitopes, but does not mean that they share a common epitope. In some examples, a partial signal by the second antibody is observed (i.e., the signal is less than the signal observed when the first antibody was not added but greater than the background), which means that the epitopes are partially overlapping.

[0082] "Antibody fragment" includes a portion of an intact antibody, preferably the antigen-binding or variable region of an intact antibody. Examples of antibody fragments include Fab, Fab’, F(ab’)2, and Fv fragments; diabodies; linear antibodies (see, e.g., U.S. Pat. No. 5,641,870, Example 2, Zapata et al. Protein Eng. 8(10):1057-1062, 1995); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0083] Papain digestion of an antibody produces two identical antigen-binding fragments called "Fab" fragments and one residual "Fc" fragment, which is designated to reflect the ability to crystallize readily. The Fab fragment consists of the entire L chain together with the variable domain domain (VH) of the H chain, as well as the first constant domain domain (C H 1). Pepsin treatment of an antibody yields a single large F(ab')2 fragment, which generally corresponds to two disulfide-linked Fab fragments with bivalent antigen-binding activity and can still cross-link to the antigen. The Fab' fragment is different from the Fab fragment in that it has several additional residues at the carboxy terminus of the C H 1 domain. Fab'-SH is the notation herein for Fab' in which the cysteine residue(s) of the constant domain has a free thiol group. The F(ab')2 antibody fragment was originally produced as a pair of Fab' fragments with a hinge cysteine in between. Other chemical linkages of antibody fragments are also known.

[0084] The term "Fc region" as used herein is used to define the C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues within the Fc region or constant region follows the EU numbering system, also called the EU index, described in Kabat et al. Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0085] "Fv" consists of a dimer of one heavy-chain variable domain and one light-chain variable domain in a close non-covalent bond. The folding of these two domains gives rise to six hypervariable loops (three loops each from the H chain and the L chain), which provide amino acid residues for antigen binding and confer antigen-binding specificity on the antibody. However, even a single variable domain (or half of the Fv containing only three Hs specific for the antigen), although the affinity may be lower than that of the whole binding site, has the ability to recognize and bind to the antigen.

[0086] "Single-chain Fv", also abbreviated as "sFv" or "scFv", is an antibody fragment containing VH and VL antibody domains linked to a single polypeptide chain. Preferably, the sFv polypeptide further contains a polypeptide linker between the VH domain and the VL domain that allows the sFv to form the desired structure for antigen binding. For considerations regarding sFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenberg and Moore eds., Springer-Verlag, New York, pp. 269-315, 1994.

[0087] The term "diabody" refers to a small antibody fragment prepared by constructing an sFv fragment (see the previous paragraph) using a short linker (about 5-10 residues) between the VH domain and the VL domain so that inter-chain rather than intra-chain V domain pairing is achieved, resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. Bispecific diabodies are heterodimers of two "crossed" sFv fragments where the VH and VL domains of two antibodies are present on different polypeptide chains. Diabodies are fully described, for example, in EP404,097, WO93 / 11161, and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448, 1993.

[0088] The "binding domain" refers to a compound or a part of a molecule that specifically binds to a target epitope, antigen, ligand, or receptor. Binding domains include, but are not limited to, antibodies (e.g., monoclonal, polyclonal, recombinant, humanized, and chimeric antibodies), antibody fragments or portions thereof (e.g., Fab fragments, Fab’2, scFv antibodies, SMIP, domain antibodies, diabodies, minibodies, scFv-Fc, affibodies, nanobodies, and VH and / or VL domains of antibodies), receptors, ligands, aptamers, and other molecules with a specified binding partner.

[0089] A "blocking" antibody or "antagonist" antibody is an antibody that inhibits or reduces the biological activity of the antigen to which it binds. Certain blocking or antagonist antibodies substantially or completely inhibit the biological activity of the antigen. In some embodiments, the activity can be tryptase enzyme activity, e.g., protease activity. In other examples, the activity can be tryptase-mediated stimulation of bronchial smooth muscle cell proliferation and / or collagen-based contraction. In other examples, the activity can be mast cell histamine release (e.g., IgE-induced histamine release and / or tryptase-induced histamine release). The antibodies of the present invention can inhibit the biological activity of tryptase by at least about 1%, about 5%, about 10%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100%.

[0090] The term "when determined by a human tryptase beta enzyme assay using synthetic peptide S-2288 as substrate" means, in the context of the claims, that the inhibitory activity is measured according to the assay described in Example 1(A)(viii)(a). Briefly, recombinant human tryptase beta 1 tetrameric active enzyme is diluted to 0.75 nM in TNH buffer (200 mM Tris, 150 mM NaCl, 0.1 mg / mL heparin, 0.01% TRITON™ X-100, pH 8.0) and combined 1:1 with anti-tryptase antibody (diluted in PBS) in a 384-well plate. The plate is incubated for 1 hour at ambient temperature with gentle stirring. The chromogenic substrate S-2288 (Chromogenix, product number 82-0852-39), or an equivalent substrate, is diluted to 1200 μM in TNH buffer and added to the plate. The final well concentrations are 400 μM S-2288, 0.25 nM recombinant human tryptase beta 1 tetramer, 66 μg / mL heparin, and 0.10 - 222 nM anti-tryptase antibody. The plate is incubated for 40 minutes at ambient temperature with gentle stirring and then read at A 405 Read at. The IC50 of the anti-tryptase antibodies is determined from the four-parameter fit of their respective curves.

[0091] The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM, some of which are further classified into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0092] The "effector function" of an antibody refers to the biological activities attributed to the Fc region of the antibody (either the Fc region of the native sequence or a variant Fc region of the amino acid sequence), and is diverse depending on the isotype of the antibody. Examples of the effector functions of an antibody include C1q binding and complement-dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, downregulation of cell surface receptors (e.g., B cell receptor), and B cell activation.

[0093] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted Ig binds to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages), enabling these cytotoxic effector cells to specifically bind to antigen-bearing target cells and then kill the target cells by cytotoxicity. The antibody "arms" the cytotoxic cells and is absolutely necessary for such killing. While NK cells, which are primary cells mediating ADCC, express only FcγRIII, monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression in hematopoietic cells is summarized in Table 3 on page 464 of Ravetch et al. Annu. Rev. Immunol. 9:457-492, 1991. To evaluate the ADCC activity of a molecule of interest, an in vitro ADCC assay as described in U.S. Patent No. 5,500,362 or 5,821,337 can be performed. Effector cells useful in such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively, or in addition, the ADCC activity of a molecule of interest can be evaluated in vivo in an animal model such as those disclosed in Clynes et al. Proc. Natl. Acad. Sci. USA 95:652-656, 1998.

[0094] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. Preferred FcRs are native sequence human FcRs. Further, preferred FcRs are those that bind to IgG antibodies (gamma receptors), and include receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternative splicing forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences that mainly differ in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic domain (see M. Daeron, Annu. Rev. Immunol. 15:203-234, 1997 for a review). FcRs are reviewed, for example, in Ravetch et al. Annu. Rev. Immunol. 9:457-492, 1991, Capel et al. Immunomethods 4:25-34, 1994, and de Haas et al. J. Lab. Clin. Med. 126:330-41, 1995. Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein. This term also includes the neonatal receptor FcRn involved in the transfer of maternal IgGs to the fetus (see, for example, Guyer et al. J. Immunol. 117:587, 1976, and Kim et al. J. Immunol. 24:249, 1994).

[0095] "Human effector cells" are leukocytes that express one or more FcRs and perform effector functions. Preferably, these cells express at least FcγRIII and perform the ADCC effector function. Examples of human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils, with PBMCs and NK cells being preferred. Effector cells can be isolated from natural sources, such as blood.

[0096] "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 the binding of the first component of the complement system (C1q) to an antibody (of an appropriate subclass) to which their cognate antigen is bound. To evaluate complement activation, for example, a CDC assay as described in Gazzano-Santoro et al. J. Immunol. Methods 202:163, 1996 can be performed.

[0097] An "epitope" is the portion of an antigen to which an antibody selectively binds. For a polypeptide antigen, a linear epitope is a peptide portion of about 4 to 15 (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12) amino acid residues. A non-linear conformational epitope can include residues of polypeptide sequences that are proximal in the three-dimensional (3D) structure of a protein. In some embodiments, an epitope includes an amino acid that is within 4 angstroms (Å) of any atom of the antibody. In some embodiments, an epitope includes an amino acid of the tryptase protomer that is within 4 Å of any atom of the partner Fab. In certain embodiments, an epitope includes an amino acid that is within 3.5 Å, 3 Å, 2.5 Å, or 2 Å of any atom of the antibody. The antibody amino acid residues that contact the antigen (i.e., the paratope) can be determined, for example, by determining the crystal structure of the antibody complexed with the antigen or by performing hydrogen / deuterium exchange.

[0098] The term "epitope is determined by an X-ray crystallography model", when used in the context of a claim, means that the atoms of a tryptase amino acid residue (e.g., a human tryptase beta 1 residue) are determined to be within 4 Å of any atom of an anti-tryptase antibody (e.g., any anti-tryptase antibody described herein, e.g., hu31A.v11 or huE104.v2) in an X-ray crystallography model such as that described in Example 3. In some embodiments, the X-ray crystallography model has a resolution of about 3.5 Å or less, about 3 Å or less, about 2.5 Å or less, about 2.15 Å or less, or about 2 Å or less.

[0099] The terms "full-length antibody", "intact antibody", and "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially the same as a native antibody structure or having a heavy chain that includes an Fc region as defined herein.

[0100] A "human antibody" is one having an amino acid sequence corresponding to that of an antibody produced by a human and / or made using any of the techniques for making human antibodies. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues.

[0101] The "human consensus framework" is a framework that represents the amino acid residues that most commonly occur in the selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is derived from a subgroup of variable domain sequences. Generally, this subgroup of sequences is a subgroup such as that in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD, vols. 1-3, 1991. In one embodiment, for VL, the subgroup is subgroup kappa III or kappa IV as in Kabat et al. (supra). In one embodiment, for VH, the subgroup is subgroup III in Kabat et al. (supra).

[0102] A "humanized" antibody of a non-human (e.g., rodent) antibody is a chimeric antibody that contains a minimal amount of sequence derived from the non-human antibody. Most often, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the hypervariable regions of the recipient are replaced by residues from the hypervariable regions of a non-human species such as a mouse, rat, rabbit, or non-human primate (donor antibody) that have the desired antibody specificity, affinity, and capacity. In some cases, framework region (FR) residues of the human immunoglobulin are replaced by the corresponding non-human residues. Additionally, a humanized antibody can contain residues not found in the recipient antibody or donor antibody. These modifications are made to further improve the antibody's capacity. Generally, a humanized antibody will contain substantially all of at least one, 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 FRs are those of a human immunoglobulin sequence. A humanized antibody optionally will also contain at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see 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.

[0103] An "immune complex" is an antibody that is complexed to one or more heterologous molecule(s) including, but not limited to, a cytotoxic agent.

[0104] When used to describe the various antibodies disclosed herein, the term "isolated" means an antibody that has been identified, separated and / or recovered from the cells or cell culture in which it is expressed. Contaminating components of its natural environment are materials that typically interfere with the diagnostic or therapeutic use of the polypeptide and can include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is purified to a purity of greater than 95% or 99% as determined by methods such as electrophoresis (e.g., sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing electrophoresis (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al. J. Chromatogr. B 848:79-87, 2007. In preferred embodiments, the antibody is purified to a sufficient extent to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a spinning cup sequenator or to homogeneity by SDS-PAGE using Coomassie blue or preferably silver staining under non-reducing or reducing conditions. Isolated antibodies include antibodies in situ within recombinant cells because at least one component of the polypeptide's natural environment is absent. However, usually, an isolated polypeptide is prepared by at least one purification step.

[0105] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical and / or bind to the same epitope on the antigen, except for possible variants that are present, for example, naturally occurring mutations or variant antibodies that occur during the production of the monoclonal antibody preparation, such variants generally being present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on the antigen. Thus, the modifier "monoclonal" indicates the characteristic of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies used in accordance with the present invention can be made by a variety of techniques including, but not limited to, the hybridoma method, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals that contain all or part of the human immunoglobulin loci, and such methods and other exemplary methods for making monoclonal antibodies are described herein. In certain embodiments, the term "monoclonal antibody" includes bispecific antibodies.

[0106] The term "bivalent antibody" refers to an antibody having two binding sites for an antigen. Bivalent antibodies can be in the IgG format or F(ab’)2 format, without limitation.

[0107] The term "multispecific antibody" is used in the broadest sense and encompasses antibodies that bind to two or more determinants or epitopes on one antigen, or two or more determinants or epitopes on two or more antigens. Such multispecific antibodies include, but are not limited to, full-length antibodies, antibodies having two or more VL and VH domains, antibody fragments such as Fab, Fv, dsFv, scFv, diabodies, bispecific diabodies, and triabodies, covalently or non-covalently linked antibody fragments. "Polyepitope specificity" refers to the ability to specifically bind to two or more different epitopes on the same or different target(s). In certain embodiments, the multispecific antibody is a bispecific antibody. "Dual specificity" or "bispecificity" refers to the ability to specifically bind to two different epitopes on the same or different target(s). However, in contrast to bispecific antibodies, dual specificity antibodies have two antigen-binding arms with identical amino acid sequences and can recognize two antigens with each Fab arm. Due to dual specificity, the antibody can interact with two different antigens with high affinity as a single Fab or IgG molecule. According to one embodiment, the multispecific antibody binds to each epitope with an affinity of 5 μM to 0.001 pM, 3 μM to 0.001 pM, 1 μM to 0.001 pM, 0.5 μM to 0.001 pM, or 0.1 μM to 0.001 pM. "Monospecificity" refers to the ability to bind to only one epitope.

[0108] A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (e.g., a cytotoxic moiety) or a radiolabel. A naked antibody may be present in a pharmaceutical composition.

[0109] With respect to the binding of an antibody to a target molecule, the terms "binds to" or "binds" or "specifically binds" or "specifically binds to" or "is specific for" a particular polypeptide or an epitope on a particular polypeptide target mean a binding that is somewhat different from non-specific interactions. Specific binding can be measured, for example, by determining the binding of a molecule relative to the binding of a control molecule. For example, specific binding can be determined by competition with a control molecule similar to the target, e.g., an excess of unlabeled target. In this case, specific binding is indicated if the binding of the labeled target to the probe is competitively inhibited by the excess of unlabeled target. The term "specific binding to" or "specifically binds to" or "is specific for" a particular polypeptide or an epitope on a particular polypeptide target, as used herein, means, for example, that the K D is, for example, 10 -4 M or less, or 10 -5 M or less, or 10 -6 M or less, or 10 -7 M or less, or 10 -8 M or less, or 10 -9 M or less, or 10 -10 M or less, or 10 -11 M or less, or 10 -12 M or less, or 10 -4 M to 10 -6 M, or 10 -6 M to 10 -10 M or 10 -7 M to 10 -9 M and can be exhibited by a molecule. As will be appreciated by those skilled in the art, affinity is inversely proportional to K D . A high affinity for an antigen is measured by a low K D value. In one embodiment, the term "specific binding" means a binding in which a molecule binds to a particular polypeptide or an epitope on a particular polypeptide without substantially binding to any other polypeptide or polypeptide epitope.

[0110] "Paratope" means a part of an antibody that binds to an epitope of an antigen. A paratope is typically a region of about 15 to 22 amino acid residues in the Fv region of an antibody and may contain amino acids from the V H and V L chains of the antibody.

[0111] The term "variable" refers to the fact that certain segments of the variable domain vary extensively between antibodies in an array. The variable or "V" domain mediates antigen binding and defines the specificity of a particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domain, which is 110 amino acids in length. Instead, the V region consists of relatively invariant ranges called framework regions (FRs) of 15-30 amino acids separated by shorter regions of extreme variability called "hypervariable regions", each 9-12 amino acids in length. The term "hypervariable region" or "HVR", as used herein, refers to the amino acid residues of the antibody involved in antigen binding. Hypervariable regions generally include, for example, residues approximately 24-34 (L1), 50-56 (L2), and 89-97 (L3) in VL, and residues approximately 26-35 (H1), 49-65 (H2), and 95-102 (H3) (in one embodiment, H1 is residues approximately 31-35), amino acid residues from Kabat et al (supra), and / or "hypervariable loops" (e.g., residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in VL, and residues 26-32 (H1), 53-55 (H2), and 96-101 (H3) in VH; Chothia et al. J. Mol. Biol. 196:901-917, 1987). The variable domains of native heavy and light chains each predominantly adopt a beta-sheet conformation connected by three hypervariable regions that form loops connecting and in some cases forming part of the beta-sheet structure, and include four FRs. The hypervariable regions in each chain are held in proximity to the hypervariable regions of the other chain by the FRs and contribute to the formation of the antigen-binding site of the antibody (see Kabat et al (supra)). Thus, HVR and FR sequences generally occur in the following sequence in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4. The constant domain does not directly participate in the binding of the antibody to the antigen, but exhibits various effector functions such as involvement in antibody-dependent cell cytotoxicity (ADCC) of the antibody.

[0112] The terms "Kabat - like variable domain residue numbering" or "Kabat - like amino acid position numbering", and variations thereof, refer to the numbering system used for the heavy - chain variable domain or the light - chain variable domain of antibody compilations in Kabat et al. (supra). Using this numbering system, the actual linear amino - acid sequence may contain fewer or additional amino acids corresponding to deletions or insertions in the FR or HVR of the variable domain. For example, the heavy - chain variable domain may contain a single amino - acid insert (residue 52a according to Kabat) after residue 52 of H2, and residues inserted after heavy - chain FR residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat, etc.). The Kabat numbering of residues can be determined for a given antibody by alignment in the homology region of the antibody's sequence with the "standard" Kabat - numbered sequence.

[0113] The Kabat numbering system is generally used when referring to residues within the variable domain (approximately residues 1 - 107 of the light chain and residues 1 - 113 of the heavy chain) (e.g., Kabat et al. (supra)). The "EU numbering system" or "EU index" is generally used when referring to residues within the immunoglobulin heavy - chain constant region (e.g., the EU index reported in Kabat et al. (supra)). "EU index like that in Kabat" refers to the residue numbering of human IgG1 EU antibodies. Unless otherwise indicated herein, references to residue numbers in the variable domain of an antibody mean residue numbering according to the Kabat numbering system. Unless otherwise indicated herein, references to residue numbers in the constant domain of an antibody mean residue numbering according to the EU numbering system (e.g., see the drawings related to EU numbering in U.S. Provisional Patent Application No. 60 / 640,323).

[0114] "Disorder" or "disease" is any condition that would benefit from treatment with an antibody (e.g., any tryptase antibody described herein). This includes chronic and acute disorders or diseases, including pathological conditions that predispose a mammal to the disorder in question. In some embodiments, the disorder is a pulmonary disorder, an immunodeficiency, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, a lymphocytic disorder, or a disorder associated with an increased number or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium). In some embodiments, the disorder is a pulmonary disorder. In some examples, the disorder can be a tryptase-related disorder or a tryptase-mediated disorder.

[0115] As used herein, the terms "tryptase-related disorder" and "tryptase-mediated disorder" refer to any disorder or condition that is mediated by or associated with tryptase. In some embodiments, a tryptase-related disorder is associated with excessive tryptase levels or activity at which atypical symptoms can occur due to local and / or systemic tryptase levels or activity in the body.

[0116] In some embodiments, the pulmonary disorder is asthma. In some embodiments, the asthma is persistent chronic severe asthma with acute episodes of worsening symptoms (aversion or sudden onset) that can be life-threatening. In some embodiments, the asthma is atopic (also known as allergic) asthma, non-allergic asthma (e.g., often induced by infections with respiratory viruses (such as influenza, parainfluenza, rhinovirus, human metapneumovirus, and respiratory syncytial virus), or inhaled irritants (such as air pollutants, smoke, diesel particles, volatile chemicals and gases indoors or outdoors, or even cold dry air)).

[0117] In some embodiments, asthma is intermittent or exercise-induced, or asthma resulting from acute or chronic direct or indirect exposure to "smoke" (typically cigarettes, cigars, pipes), inhalation or "vaping" (tobacco, marijuana, or other such substances), or asthma induced by recent ingestion of aspirin or related NSAIDs. In some embodiments, asthma is mild or corticosteroid-naive asthma, newly diagnosed untreated asthma, or asthma that has not previously required chronic use of topical or systemic inhaled steroids to suppress symptoms (cough, wheeze, shortness of breath / dyspnea, or chest pain). In some embodiments, asthma is chronic, corticosteroid-resistant asthma, corticosteroid-refractory asthma, asthma that is not controlled by corticosteroids or other chronic asthma controller medications.

[0118] In some embodiments, the asthma is moderate to severe asthma. In certain embodiments, the asthma is Th2-high asthma. In some embodiments, the asthma is severe asthma. In some embodiments, the asthma is allergic asthma, allergic asthma, non-allergic asthma (e.g., due to infections and / or respiratory syncytial virus (RSV)), exercise-induced asthma, aspirin-sensitive / intolerant asthma, mild asthma, moderate to severe asthma, corticosteroid-naive asthma, chronic asthma, corticosteroid-resistant asthma, corticosteroid-refractory asthma, newly diagnosed untreated asthma, smoking-induced asthma, asthma uncontrolled by corticosteroids. In some embodiments, the asthma is type 2 helper T lymphocyte (Th2) or type 2 (Th2)-high helper T lymphocyte, or type 2 (T2)-driven asthma. In some embodiments, the asthma is eosinophilic asthma. In some embodiments, the asthma is allergic asthma. In some embodiments, the individual has been determined to be eosinophilic inflammation positive (EIP). See WO2015 / 061441. In some embodiments, the asthma is periostin-high asthma (e.g., having a periostin level of at least about 20 ng / mL, 25 ng / mL, or 50 ng / mL of serum). In some embodiments, the asthma is eosinophil-high asthma (e.g., any of at least about 150, 200, 250, 300, 350, 400 eosinophils / ml of blood). In certain embodiments, the asthma is Th2-low asthma or non-Th2-driven asthma. In some embodiments, the individual has been determined to be eosinophilic inflammation negative (EIN). See WO2015 / 061441. In some embodiments, the asthma is periostin-low asthma (e.g., having a periostin level of less than about 20 ng / mL of serum). In some embodiments, the asthma is eosinophil-low asthma (e.g., less than about 150 eosinophils / μl of blood or less than about 100 eosinophils / μl of blood).

[0119] As used herein, the term "Th2-high asthma" refers to asthma presenting high levels of one or more Th2 cell-related cytokines such as IL13, IL4, IL9, IL5, or presenting Th2 cytokine-related inflammation. In certain embodiments, the term "Th2-high asthma" may be used interchangeably with eosinophilic-high asthma. In certain embodiments, Th2-high asthma is Th2-driven asthma. In some embodiments, an asthma patient has been determined to be eosinophilic inflammation positive (EIP). See, e.g., International Patent Application Publication No. WO2015 / 061441, which is incorporated herein by reference in its entirety. In certain embodiments, an individual has been determined to have an elevated level of at least one of the eosinophilic signature genes as compared to a control or reference level. See WO2015 / 061441. In certain embodiments, Th2-high asthma is periostin-high asthma. In some embodiments, an individual has high serum periostin. In certain embodiments, an individual is 18 years of age or older. In certain embodiments, an individual has been determined to have an elevated level of serum periostin as compared to a control or reference level. In certain embodiments, the control or reference level is the median level of periostin in the population. In certain embodiments, an individual has been determined to have serum periostin of 20 ng / ml or more. In certain embodiments, an individual has been determined to have serum periostin of 25 ng / ml or more. In certain embodiments, an individual has been determined to have serum periostin of 50 ng / ml or more. In certain embodiments, the control or reference level of serum periostin is 20 ng / ml, 25 ng / ml, or 50 ng / ml. In certain embodiments, the asthma is eosinophilic-high asthma. In certain embodiments, an individual has been determined to have an elevated eosinophil count as compared to a control or reference level. In certain embodiments, the control or reference level is the median level of the population. In certain embodiments, an individual has been determined to have an eosinophil count of 150 or more per μl of blood. In certain embodiments, an individual has been determined to have an eosinophil count of 200 or more per μl of blood.In certain embodiments, the individual is determined to have an eosinophil count of 250 or more eosinophils / μl of blood. In certain embodiments, the individual is determined to have an eosinophil count of 300 or more eosinophils / μl of blood. In certain embodiments, the individual is determined to have an eosinophil count of 350 or more eosinophils / μl of blood. In certain embodiments, the individual is determined to have an eosinophil count of 400 or more eosinophils / μl of blood. In certain embodiments, the individual is determined to have an eosinophil count of 450 or more eosinophils / μl of blood. In certain embodiments, the individual is determined to have an eosinophil count of 500 or more eosinophils / μl of blood. In certain preferred embodiments, the individual is determined to have an eosinophil count of 300 or more eosinophils / μl of blood. In certain embodiments, the eosinophils are peripheral blood eosinophils. In certain embodiments, the eosinophils are sputum eosinophils. In certain embodiments, the individual exhibits elevated levels of FeNO (fractional exhaled nitric oxide) and / or elevated levels of IgE. For example, in some instances, the individual exhibits an FeNO level of greater than about 250 parts per billion (ppb), greater than about 275 ppb, greater than about 300 ppb, greater than about 325 ppb, greater than about 325 ppb, or greater than about 350 ppb. In some instances, the individual has an IgE level of greater than 50 IU / ml.

[0120] As used herein, the terms "Th2-low asthma" or "non-Th2-high asthma" refer to asthma presenting low levels of one or more Th2 cell-related cytokines, such as IL13, IL4, IL9, IL5, or presenting non-Th2 cytokine-related inflammation. In certain embodiments, the term Th2-low asthma may be used interchangeably with eosinophilic-low asthma. In some embodiments, asthma patients have been determined to be eosinophilic inflammation negative (EIN). See, e.g., WO2015 / 061441. In certain embodiments, Th2-low asthma is Th17-driven asthma. In certain embodiments, Th2-low asthma is periostin-low asthma. In certain embodiments, the individual is 18 years of age or older. In certain embodiments, the individual has been determined to have a reduced level of serum periostin as compared to a control or reference level. In certain embodiments, the control or reference level is the median level of periostin in the population. In certain embodiments, the individual has been determined to have serum periostin of less than 20 ng / ml. In certain embodiments, the asthma is eosinophilic-low asthma. In certain embodiments, the individual has been determined to have a reduced number of eosinophils as compared to a control or reference level. In certain embodiments, the control or reference level is the median level of the population. In certain embodiments, the individual has been determined to have less than 150 eosinophils / μl blood. In certain embodiments, the individual has been determined to have less than 100 eosinophils / μl blood. In certain preferred embodiments, the individual has been determined to have less than 300 eosinophils / μl blood.

[0121] In some embodiments, an autoimmune disorder, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, or a lymphocytic disorder is esophagitis (e.g., eosinophilic esophagitis), allergic rhinitis, non-allergic rhinitis, rhinosinusitis with polyps, nasal polyps, bronchitis, chronic pneumonia, allergic bronchopulmonary aspergillosis, airway inflammation, allergic rhinitis, bronchiectasis, and / or chronic bronchitis.

[0122] In some embodiments, the autoimmune deficiency, inflammatory disorder, fibrotic disorder, granulocytic (neutrophilic or eosinophilic) disorder, monocytic disorder, or lymphocytic disorder is arthritis. In some embodiments, the arthritis is rheumatoid arthritis. In some embodiments, the arthritis is osteoarthritis, rheumatoid arthritis, juvenile arthritis, juvenile rheumatoid arthritis, early arthritis, polyarticular rheumatoid arthritis, systemic rheumatoid arthritis, enteropathic arthritis, reactive arthritis, psoriatic arthritis, and / or arthritis as a result of an injury.

[0123] In some embodiments, the autoimmune deficiency, inflammatory disorder, fibrotic disorder, granulocytic (neutrophilic or eosinophilic) disorder, monocytic disorder, or lymphocytic disorder is a gastroenteritis condition. In some embodiments, the gastroenteritis condition is IBD (inflammatory bowel disease), ulcerative colitis (UC), Crohn's disease (CD), colitis (e.g., colitis caused by external stimuli (e.g., caused by or associated with a treatment regimen, e.g., chemotherapy, radiation therapy, etc.)), infectious colitis, ischemic colitis, collagenous or lymphocytic colitis, necrotizing enteritis, colitis in conditions such as chronic granulomatous disease or celiac disease, food allergy, gastritis, gastroenteritis, infectious gastritis or enteritis (e.g., Helicobacter pylori infection chronic active gastritis), esophagitis, and other forms of gastrointestinal inflammation caused by an infective agent, or ulcerative colitis.

[0124] In some embodiments, an autoimmune disorder, an inflammatory disorder, a fibrotic disorder, a granulocytic (neutrophilic or eosinophilic) disorder, a monocytic disorder, or a lymphocytic disorder is a gastroenteritis condition. In some embodiments, the gastroenteritis condition is IBD (inflammatory bowel disease). In some embodiments, the inflammatory bowel disease is ulcerative colitis (UC) or Crohn's disease (CD). In some embodiments, the gastroenteritis condition is colitis (e.g., colitis caused by external stimuli (e.g., caused by or associated with a treatment regimen, e.g., chemotherapy, radiation therapy, etc.)), infectious colitis, ischemic colitis, collagenous or lymphocytic colitis, necrotizing enteritis, colitis in conditions such as chronic granulomatous disease or celiac disease, food allergy, gastritis, gastroenteritis, infectious gastritis or enteritis (e.g., Helicobacter pylori - infected chronic active gastritis), and other forms of gastrointestinal inflammation caused by an infectious agent, or ulcerative colitis. In some embodiments, the gastroenteritis condition is ulcerative colitis (UC) or Crohn's disease (CD). In some embodiments, the gastroenteritis condition is ulcerative colitis (UC). In some embodiments, ulcerative colitis is mild to moderate distal colitis. In some embodiments, ulcerative colitis is mild to moderate extensive colitis. In some embodiments, ulcerative colitis is severe colitis. In some embodiments, the gastroenteritis condition is Crohn's disease (CD). In some embodiments, Crohn's disease is in an acute disease phase. In some embodiments, Crohn's disease is in an induction clinical remission phase. In some embodiments, Crohn's disease is in a sustained response / remission phase. In some embodiments, Crohn's disease is a mild to moderate disease. In some embodiments, Crohn's disease is a moderate to severe disease. In some embodiments, Crohn's disease is a severe / critical disease. In some embodiments, Crohn's disease is an ileal, ileocolonic, or colonic disease.

[0125] In some embodiments, disorders associated with increased number or distribution of autoimmune deficiencies, inflammatory disorders, fibrotic disorders, granulocytic (neutrophilic or eosinophilic) disorders, monocytic disorders, or lymphocytic disorders, or normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium) are one or more organ-specific symptoms of lupus or systemic lupus erythematosus (SLE), or lupus (e.g., lupus nephritis (LN) affecting the kidney, or extrarenal lupus (ERL) affecting the blood and / or lymphatic organs (lymph nodes, spleen, thymus, and associated lymphatics), and / or joints and / or other organs, but not necessarily the kidney).

[0126] In some embodiments, the autoimmune disorder, inflammatory disorder, or fibrotic disorder is associated with sepsis and / or trauma, HIV infection, or idiopathic (unknown etiology) multiple vasculitic granulomatosis (previously known as Wegener's granulomatosis), Behçet's disease, cardiovascular disease, eosinophilic bronchitis, Reiter's syndrome, SEA syndrome (seronegative, enthesopathy, arthritis syndrome), ankylosing spondylitis, dermatomyositis, scleroderma, for example, systemic scleroderma also known as systemic sclerosis, vasculitis (e.g., temporal arteritis, cranial arteritis, or giant cell arteritis (GCA) also known as Horton's disease), myositis, polymyositis, dermatomyositis, arteritis, rheumatoid polymyalgia, sarcoidosis, primary biliary cirrhosis, sclerosing cholangitis, Sjögren's syndrome, psoriasis, plaque psoriasis, guttate psoriasis, inverse psoriasis, pustular psoriasis, psoriatic erythroderma, dermatitis, atopic dermatitis, pemphigus, for example, pemphigus vulgaris, atherosclerosis, lupus, Still's disease, myasthenia gravis, celiac disease, relapsing remitting (RRMS) or primary progressive (PPMS) or secondary progressive (SPMS) subtype of multiple sclerosis (MS), Guillain - Barré disease, type I diabetes (T1DM) or insulin - dependent (IDDM) or juvenile - onset DM type, thyroiditis (e.g., Graves' disease), celiac disease, Churg - Strauss syndrome, myalgic syndrome, eosinophilic syndrome, an edema reaction including eosinophilic angioedema, helminth infection, onchocerciasis dermatitis, eosinophilic esophagitis, eosinophilic enteritis, eosinophilic colitis, obstructive sleep apnea, endomyocardial fibrosis, Addison's disease, Raynaud's disease or phenomenon, autoimmune hepatitis, graft - versus - host disease (GVHD), or organ transplant rejection.

[0127] In some embodiments, the disorder is an inflammatory disorder of the skin. In some embodiments, the disorder is atopic dermatitis or onchocerciasis dermatitis. In some embodiments, the disorder is chronic idiopathic urticaria (CIU or CSU).

[0128] In some embodiments, the autoimmune disorder, inflammatory disorder, fibrotic disorder, neutrophil disorder, or eosinophilic disorder is a fibrotic disorder. In some embodiments, the fibrotic disorder is pulmonary fibrosis, liver fibrosis (e.g., fibrosis associated with cirrhosis (e.g., alcohol-induced cirrhosis, virus-induced cirrhosis, post-hepatitis C cirrhosis, and primary biliary cirrhosis), schistosomiasis, cholangitis (e.g., sclerosing cholangitis), and autoimmune-induced hepatitis), kidney fibrosis (e.g., tubulointerstitial fibrosis, scleroderma, diabetic nephropathy, and glomerulonephritis), skin fibrosis (e.g., scleroderma, hypertrophic and keloid scars, nephrogenic fibrosing dermopathy, and burns), myelofibrosis, neurofibromatosis, fibroma, intestinal fibrosis, and fibrotic adhesions resulting from surgical procedures), cardiac fibrosis (e.g., fibrosis associated with myocardial infarction), vascular fibrosis (e.g., fibrosis associated with post-angioplasty arterial restenosis and atherosclerosis), ocular fibrosis (e.g., fibrosis associated with proliferative vitreoretinopathy after cataract surgery and retro-orbital fibrosis), and myelofibrosis (e.g., idiopathic myelofibrosis and drug-induced myelofibrosis). The fibrosis can be organ-specific or systemic (e.g., systemic sclerosis and fibrosis associated with GVHD). In some embodiments, the fibrotic disorder is pulmonary fibrosis. In some embodiments, the pulmonary fibrosis is fibrotic interstitial pneumonia. In some embodiments, the pulmonary fibrosis is idiopathic pulmonary fibrosis (IPF), also known as idiopathic fibrosing alveolitis. In some embodiments, the IPF is gender, age, and physiology (GAP) stage I. In some embodiments, the IPF is GAP stage II. In some embodiments, the IPF is GAP stage III. In some embodiments, the pulmonary fibrosis is sporadic IPF. In some embodiments, the pulmonary fibrosis is familial pulmonary fibrosis. In some embodiments, the pulmonary fibrosis is combined pulmonary fibrosis and emphysema.In some embodiments, pulmonary fibrosis is associated with one or more of the following: usual interstitial pneumonia, idiopathic interstitial pneumonia, desquamative interstitial pneumonia, respiratory bronchiolitis interstitial lung disease, acute interstitial pneumonia, nonspecific interstitial pneumonia, sarcoidosis, idiopathic pulmonary fibrosis, eosinophilic pneumonia, infectious diseases, occupational or environmental substance exposure, cigarette smoking, drug- or radiation-induced interstitial lung disease, rheumatic disease-related interstitial lung disease, lymphocytic interstitial pneumonia, upper lobe predominant pulmonary fibrosis, pulmonary Langerhans cell histiocytosis, systemic sclerosis interstitial lung disease, Hermansky-Pudlak syndrome, and diseases due to telomere shortening.

[0129] In some embodiments, a lung disorder, an autoimmune disorder, an inflammatory disorder, a fibrotic disorder, a neutrophilic disorder, or an eosinophilic disorder is chronic obstructive pulmonary disease (COPD). In some embodiments, COPD is the international guideline regarding chronic obstructive pulmonary disease (GOLD) category A. In some embodiments, COPD is GOLD category B. In some embodiments, COPD is GOLD category C. In some embodiments, COPD is GOLD category D. In some embodiments, COPD is chronic bronchitis. In some embodiments, COPD is emphysema. In some embodiments, emphysema is centrilobular, panlobular, or paraseptal emphysema. In some embodiments, emphysema is tobacco-induced emphysema. In some embodiments, COPD is associated with exposure to particulate dust, chemical fumes, and / or air pollution. In some embodiments, COPD is associated with a disorder of lung development. In some embodiments, COPD is chronic obstructive asthma. In some embodiments, COPD is associated with alpha1-antitrypsin deficiency. In some embodiments, COPD is associated with the cleavage of serine protease inhibitor clade E, member 2 (SERPINE2). In some embodiments, COPD is COPD with persistent systemic inflammation. In some embodiments, COPD is eosinophilic or type 2 helper T (T H2) It is severe COPD. In some embodiments, the COPD is COPD with persistent bacterial colonization. In some embodiments, the COPD is COPD with frequent exacerbations. In some embodiments, the autoimmune disorder, inflammatory disorder, fibrotic disorder, neutrophilic disorder, or eosinophilic disorder is asthmatic COPD overlap syndrome (ACOS). In some embodiments, the ACOS is eosinophil-predominant, neutrophil-predominant, mixed pattern, or non-inflammatory (non-granulocytic) ACOS. In some embodiments, the autoimmune disorder, inflammatory disorder, fibrotic disorder, neutrophilic disorder, or eosinophilic disorder is COPD obstructive sleep apnea (OSA) overlap syndrome.

[0130] The above list is not comprehensive, and those skilled in the art will understand that a disease or disorder can fall within various categories.

[0131] The term "package insert" is used to refer to the instructions customarily included in the commercial package of a therapeutic product that contains information about the indications, usage, dosage, administration, combination therapy, contraindications, and / or warnings regarding its use.

[0132] The terms "pharmaceutical formulation" and "pharmaceutical composition" are used interchangeably herein and refer to a preparation that is in a form in which the biological activity of the active ingredient contained therein is effective and that does not contain additional ingredients that are toxic to the extent that the preparation is unacceptable to the subject to which the formulation is administered. Such a formulation is sterile. In a preferred embodiment, the pharmaceutical composition or pharmaceutical formulation is administered to a human subject.

[0133] A "sterile" pharmaceutical formulation is aseptic or does not contain, or essentially does not contain, all viable microorganisms and their spores.

[0134] A "stable" pharmaceutical formulation is a formulation in which the internal protein (e.g., an antibody such as an anti - tryptase antibody) essentially retains its physical stability and / or chemical stability and / or biological activity during storage. Preferably, the formulation essentially retains its physical and chemical stability, as well as its biological activity, during storage. The storage period is generally selected based on the intended shelf - life of the formulation. Various analytical techniques for measuring protein stability are available in the art and are outlined, for example, in Peptide and Protein Drug Delivery, 247 - 301, Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991), Jones, A. Adv. Drug Delivery Rev. 10:29 - 90 (1993). Stability can be measured over a selected period at a selected exposure and / or temperature. Stability can be qualitatively and / or quantitatively evaluated in a variety of different ways, including assessment of aggregate formation (e.g., by measuring turbidity using size - exclusion chromatography and / or by visual inspection); assessment of ROS formation (e.g., by using a photo - stress assay or an AAPH stress assay); oxidation of specific amino acid residues of the protein (e.g., Trp residues and / or Met residues of a monoclonal antibody); assessment of charge heterogeneity using cation - exchange chromatography, imaging capillary isoelectric focusing (icIEF), or capillary zone electrophoresis; amino - terminal or carboxy - terminal sequence analysis; mass spectrometry; SDS - PAGE analysis for comparing reduced intact antibodies; peptide mapping (e.g., trypsin or LYS - C) analysis; assessment of the biological activity or target - binding function of the protein (e.g., the antigen - binding function of an antibody), etc. Instability can be associated with any one or more of aggregation, deamidation (e.g., Asn deamidation), oxidation (e.g., Met oxidation and / or Trp oxidation), isomerization (e.g., Asp isomerization), clipping / hydrolysis / fragmentation (e.g., hinge - region fragmentation), succinimide formation, unpaired cysteine(s), N - terminal extension, C - terminal processing, glycosylation differences, etc.

[0135] An antibody (e.g., an anti - tryptase antibody) "retains its physical stability" in a pharmaceutical formulation if it shows little or no sign of aggregation, precipitation, fragmentation, and / or denaturation upon visual inspection of color and / or transparency or when measured by UV light scattering or size - exclusion chromatography.

[0136] An antibody (e.g., an anti - tryptase antibody) "retains its chemical stability" in a pharmaceutical formulation if its chemical stability at a given time point is such that the antibody is still considered to retain its biological activity as defined below. Chemical stability can be assessed by detecting and quantifying chemically modified forms of the antibody. Chemical modifications can include, for example, protein oxidation that can be evaluated using trypsin peptide mapping, reverse - phase high - performance liquid chromatography (HPLC), and liquid chromatography - mass spectrometry (LC / MS). Other types of chemical modifications can include, for example, charge modifications of the antibody that can be evaluated by ion - exchange chromatography or icIEF.

[0137] An antibody (e.g., an anti - tryptase antibody) "retains its biological activity" in a pharmaceutical formulation if its biological activity at a given time point is within about 20% (e.g., within about 10%) of the biological activity exhibited at the time the pharmaceutical formulation was prepared, as determined, for example, by an antigen - binding assay or an in vitro inhibition assay of a monoclonal antibody (e.g., an anti - tryptase monoclonal antibody) (within the error of the assay). In some embodiments, the biological activity of the antibody at a given time point is about 25%, about 30%, about 35%, about 40%, about 45%, about 50% of the biological activity exhibited at the time the pharmaceutical formulation was prepared.

[0138] As used herein, the "biological activity" of an antibody (e.g., an anti-tryptase antibody) refers to the ability of the antibody to bind to a target, e.g., the ability of a monoclonal antibody to bind to an antigen. This may further include a biological response that can be measured in vitro or in vivo. Such activity can be antagonist activity or agonist activity.

[0139] An "oxidation-sensitive" protein (e.g., an antibody such as an anti-tryptase antibody) is a protein that contains one or more residues (s) that have been found to be prone to oxidation, such as, but not limited to, methionine (Met), cysteine (Cys), histidine (His), tryptophan (Trp), and tyrosine (Tyr). For example, a tryptophan amino acid within the Fab portion of a monoclonal antibody or a methionine amino acid within the Fc portion of a monoclonal antibody can be oxidation-sensitive.

[0140] The term "percent oxidation" refers to the percentage of an antibody in a formulation (e.g., a pharmaceutical composition) that is oxidized at a particular amino acid residue, e.g., a Trp residue (e.g., Trp100 in HVR-H3 of hu31A.v11) or a Met residue. The percent oxidation can be determined, for example, by mass spectrometry (MS) of one or more tryptic peptides in which one or more specific oxidized proline amino acid residues are present. In certain embodiments, the percent oxidation of Trp100 in HVR-H3 of hu31A.v11 is determined by the mass of the oxidized tryptic peptide in which Trp100 is present in excess of the mass of the total (oxidized and non-oxidized) tryptic peptide as determined by MS analysis. The percent oxidation can be determined, for example, within 9 months, 12 months, 18 months, or 2 years from the initial production of the antibody or its pharmaceutical composition.

[0141] As used herein, the term "determined according to the AAPH stress test" means that the percentage of oxidation at a particular amino acid residue (e.g., Trp100 in HVR-H3 of hu31A.v11) is determined by mass spectrometry of tryptic peptides after formulating the antibody at 150 mg / ml in 5 mM AAPH for 25 hours at 40°C as described, for example, in Example 5. The stressed antibody was digested with trypsin and the digested peptides were subjected to ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) to determine the percentage of oxidation.

[0142] As used herein, "buffer" refers to a buffer solution that resists changes in pH due to the action of its acid-base conjugate components. The buffer of this invention preferably has a pH in the range of about 4.5 to about 8.0 (e.g., about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, or about 8), e.g., about pH 5.5. For example, histidine acetate is an example of a buffer that controls the pH within this range. Another suitable buffer is arginine succinate and / or histidine succinate.

[0143] "Preservative" is, for example, a compound that can optionally be included in a formulation to substantially reduce internal bacterial action and thus facilitate the production of multi-purpose formulations. Examples of possible preservatives include, for example, octadecyl dimethyl benzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkyl benzyl dimethyl ammonium chlorides where the alkyl group is a long-chain compound), and benzetonium chloride. Other types of preservatives include aromatic alcohols such as phenol, butyl, and benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol. In one embodiment, the preservative herein is benzyl alcohol.

[0144] As used herein, "surfactant" refers to a surface-active agent, preferably a nonionic surfactant. Examples of surfactants herein include polysorbates (e.g., polysorbate 20 and polysorbate 80), poloxamers (e.g., poloxamer 188), TRITON (registered trademark), sodium dodecyl sulfate (SDS), sodium lauryl sulfate, sodium octyl glucoside, lauryl sulfobetaine, myristyl sulfobetaine, linoleyl sulfobetaine, or stearyl sulfobetaine, lauryl sarcosine, myristyl sarcosine, linoleyl sarcosine, or stearyl sarcosine, linoleyl betaine, myristyl betaine, or cetyl betaine, lauroamidopropyl betaine, cocamidopropyl betaine, linoleamidopropyl betaine, myristamidopropyl betaine, palmidopropyl betaine, or isostearamidopropyl betaine (e.g., lauroamidopropyl), myristamidopropyldimethylamine, palmidopropyldimethylamine, or isostearamidopropyldimethylamine, sodium methyl cocoyl taurate or disodium methyl oleoyl taurate, and MONAQUAT (trademark) series (Mona Industries, Inc., Paterson, N.J.), polyethylene glycol, polypropylene glycol, and copolymers of ethylene glycol and propylene glycol (e.g., PLURONIC (registered trademark) block copolymer type, e.g., PLURONIC (registered trademark) F-68), etc. In one embodiment, the surfactant herein is polysorbate 20. In yet another embodiment, the surfactant herein is poloxamer 188.

[0145] "Pharmaceutically acceptable carrier" refers to a component in a pharmaceutical formulation other than the active ingredient that is non-toxic to the subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0146] As used herein, the term "prodrug" refers to a precursor or derivative form of a pharmaceutically active substance that is less cytotoxic to tumor cells compared to the parent drug and can be activated by an enzyme or converted into a more active parent form. See, for example, Wilman, "Prodrugs in Cancer Chemotherapy" Biochemical Society Transactions, 14, pp. 375-382, 615th Meeting Belfast (1986) and Stella et al. "Prodrugs: A Chemical Approach to Targeted Drug Delivery," Directed Drug Delivery, Borchardt et al. (ed.), pp. 247-267, Humana Press (1985). Prodrugs of the present invention include, but are not limited to, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate-containing prodrugs, peptide-containing prodrugs, D-amino acid-modified prodrugs, glycosylated prodrugs, β-lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs or optionally substituted phenylacetamide-containing prodrugs, 5-fluorocytosine and other 5-fluorouridine prodrugs, which can be converted into a more active cytotoxic free drug. Examples of cytotoxic drugs that can be derivatized into prodrug forms for use in the present invention include, but are not limited to, the above-mentioned chemotherapeutic agents.

[0147] A "subject" is a vertebrate, preferably a mammal, more preferably a human. Mammals include, but are not limited to, domestic animals (such as cows and sheep), sporting animals, pets (such as cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys (e.g., cynomolgus monkeys)), and rodents (e.g., mice and rats).

[0148] As used herein, "administering" means a method of giving a dosage of a compound (e.g., an anti - tryptase antibody of the present invention or a further therapeutic agent) or a composition (e.g., a pharmaceutical composition, e.g., a pharmaceutical composition containing an anti - tryptase antibody of the present invention and optionally a further therapeutic agent, which may contain excipients such as antioxidants (e.g., N - acetyltryptophan and / or methionine)). The compositions used in the methods described herein can be administered, for example, intravitreally, intramuscularly, intravenously, intradermally, percutaneously, intra - arterially, intraperitoneally, intralesionally, intracranially, intra - articularly, intraprostatically, intrapleurally, intratracheally, intrathecally, intranasally, intravaginally, rectally, topically, intratumorally, intraperitoneally, subcutaneously, subconjunctivally, intravesicularly, transmucosally, epicardially, intra - umbilically, intraocularly, periorbitally, conjunctivally, sub - Tenon's capsule, intra - anterior chamber, sub - retinal, retro - ocular, intracanalicularly, by inhalation, injection, implantation, infusion, continuous infusion, local perfusion directly into target cells, catheter, lavage, cream, or lipid composition. The compositions utilized in the methods described herein can also be administered systemically or locally. The method of administration can vary depending on various factors (e.g., the compound or composition being administered and the severity of the condition, disease, or disorder being treated).

[0149] An "effective amount" or "therapeutically effective amount" of an agent, e.g., an anti - tryptase antibody or a pharmaceutical formulation (e.g., a pharmaceutical formulation containing an anti - tryptase antibody which may contain excipients such as antioxidants (N - acetyltryptophan and / or methionine)), refers to an amount effective to achieve a desired therapeutic or prophylactic result over a required period of time at a required dosage. A therapeutically effective amount of an antibody or antibody fragment (e.g., an anti - tryptase antibody), or a composition thereof, can effect a remission or treatment of a disorder or disease, or a prevention, reduction, remission, or treatment of symptoms associated with the disorder or disease.

[0150] As used herein, "treatment" (and grammatical variations thereof such as "treating" or "to treat") refers to a clinical intervention undertaken with the purpose of altering the natural course of an individual being treated and can be performed prophylactically or during a clinical pathologic process. Desirable effects of treatment include, but are not limited to, prevention of the development or recurrence of a disease, alleviation of symptoms, diminution of any direct or indirect pathologic consequences of the disease, prevention of metastasis, decrease in the rate of disease progression, regression or remission of the disease state, and improvement of palliation or prognosis. In some embodiments, the antibodies of the invention are used to delay the development of a disease or slow the progression of a disease. A patient may be considered to have had a "treatment" of asthma that was successful if, for example, after receiving a treatment regimen for asthma, the patient demonstrates a reduction or absence that is observable and / or measurable in one or more of the following: recurrent wheezing, coughing, dyspnea, chest tightness, symptoms that occur or worsen at night, symptoms that are induced by cold air, exercise, or exposure to an allergen.

[0151] The term "interleukin-5 (IL-5)" as used herein refers to any native IL-5 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. The term encompasses "full-length", unprocessed IL-5, mature IL-5, and any form of IL-5 resulting from post-translational modifications. The term also encompasses naturally occurring variants of IL-5, such as splice variants or allelic variants. An exemplary amino acid sequence of IL-5 can be found, for example, as UniProtKB accession number P05113.

[0152] The term "IL-5 axis-binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from the interaction of IL-5 with one or more of its binding partners such as interleukin-5 receptor alpha (IL5RA). Exemplary IL-5 axis-binding antagonists that can be used in the methods of the present invention include, for example, IL-5 binding antagonists (e.g., anti-IL-5 antibodies (e.g., mepolizumab, benralizumab, and reslizumab) and anti-IL-5 receptor binding antagonists (e.g., anti-IL-5R antibodies)).

[0153] As used herein, the term "interleukin-13 (IL-13)" refers to any native IL-13 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. IL-13 is a cytokine secreted by many cell types, including type 2 T helper (Th2) cells. The term encompasses "full-length", unprocessed IL-13, mature IL-13, and any form of IL-13 resulting from post-translational modification. An exemplary amino acid sequence of human IL-13 can be found, for example, as UniProtKB accession number P35225.

[0154] The term "IL-13 axis-binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from the interaction of IL-13 with one or more of its binding partners, such as interleukin-4 receptor alpha (IL4Rα), interleukin-13 receptor alpha 1 (IL13RA1), and interleukin-13 receptor alpha 2 (IL13RA2). Examples of IL-13 axis-binding antagonists include IL-13 binding antagonists (e.g., anti-IL-13 antibodies such as lebrikizumab, 228B / C-1, 228A-4, 227-26, and 227-43 (see, e.g., U.S. Patent Nos. 7,674,459; 8,067,199; 8,088,618; 8,318,160; and 8,734,797)) and IL-13 receptor binding antagonists (e.g., anti-IL4Rα antibodies, anti-IL13RA1 antibodies, or anti-IL13RA2 antibodies).

[0155] As used herein, the term "interleukin-17 (IL-17)" refers to any native IL-17 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified, and includes the family members IL-17A, IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F. The term encompasses "full-length", unprocessed IL-17, mature IL-17, and any form of IL-17 resulting from post-translational modification. The amino acid sequence of exemplary human IL-17A can be found, for example, as UniProtKB accession number Q16552. The amino acid sequence of exemplary human IL-17B can be found, for example, as UniProtKB accession number Q9UHF5. The amino acid sequence of exemplary human IL-17C can be found, for example, as UniProtKB accession number Q9P0M4. The amino acid sequence of exemplary human IL-17D can be found, for example, as UniProtKB accession number Q8TAD2. The amino acid sequence of exemplary human IL-17E can be found, for example, as UniProtKB accession number Q9H293. The amino acid sequence of exemplary human IL-17F can be found, for example, as UniProtKB accession number Q96PD4.

[0156] The term "IL-17 axis-binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from the interaction of IL-17 with one or more of its binding partners, which are interleukin-17 receptor (IL-17R) family member proteins, such as interleukin 17 receptor A (IL17RA), interleukin 17 receptor B (IL17RB), interleukin 17 receptor C (IL17RC), interleukin 17 receptor D (IL17RD), interleukin 17 receptor E (IL17RE), and interleukin 17 receptor E-like (IL17REL). Exemplary IL-17 axis-binding antagonists include, for example, IL-17 binding antagonists (e.g., anti-IL-17 antibodies (e.g., secukinumab (AIN417), ixekizumab (LY2439821), bimekizumab, and NI-1401)), and IL-17 receptor binding antagonists (e.g., anti-IL-17R antibodies (e.g., brodalumab (AMG-827))). See, for example, WO2006 / 013107, WO2007 / 070750, WO2012 / 156219, and U.S. Patent No. 8,715,669.

[0157] As used herein, the term "interleukin-33 (IL-33)" refers to any native IL-33 derived from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. IL-33 is also referred to in the art as nuclear factor of high endothelial venules (NF-HEV, see, e.g., Baekkevold et al. Am. J. Pathol. 163(1):69-79, 2003), DVS27, C9orf26, and interleukin-1 family member 11 (IL-1F11). The term encompasses "full-length", unprocessed IL-33, mature IL-33, and any form of IL-33 resulting from post-translational modification. Human full-length, unprocessed IL-33 contains 270 amino acids (a.a.) and IL-33 1-270which may also be referred to as. Processing forms of human IL-33 include, for example, IL-33 95-270 , IL-33 99-270 , IL-33 109-270 , IL-33 112-270 , IL-33 1-178 , and IL-33 179-270 (Lefrancais et al. Proc. Natl. Acad. Sci. 109(5):1673-1678, 2012 and Martin, Semin. Immunol. 25:449-457, 2013). In some embodiments, processing forms of human IL-33, such as IL-33 95-270 , IL-33 99-270 , IL-33 109-270 , or other forms processed by proteases such as calpain, proteinase 3, neutrophil elastase, and cathepsin G may have increased biological activity compared to full-length IL-33. The term also encompasses naturally occurring variants of IL-33, such as splice variants (e.g., the constitutively active splice variant spIL-33 lacking exon 3, Hong et al. J. Biol. Chem. 286(22):20078-20086, 2011) or allelic variants. IL-33 may be present intracellularly (e.g., in the nucleus) or in secreted cytokine form. The full-length IL-33 protein contains a helix-turn-helix DNA-binding motif containing a nuclear localization sequence (amino acids 1-75 of human IL-33), which includes a chromatin-binding motif (amino acids 40-58 of human IL-33). The processed and secreted forms of IL-33 lack these N-terminal motifs. An exemplary amino acid sequence of human IL-33 can be found, for example, as UniProtKB accession number O95760.

[0158] As used interchangeably herein, the terms "interleukin 1 receptor-like 1 (IL1RL1)" and "ST2" refer to any native ST2 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. ST2 is also referred to in the art as DER4, T1, and FIT-1. The term encompasses "full-length", unprocessed ST2, mature ST2, and any form of ST2 resulting from post-translational modifications. At least four isoforms of ST2 are known in the art, including soluble (also known as IL1RL1-a, sST2) and transmembrane (ST2L, also known as IL1RL1-b) resulting from differential mRNA expression from a dual promoter system, and ST2V and ST2LV resulting from alternative splicing, as described below. The domain structure of ST2L includes three extracellular immunoglobulin-like C2 domains, a transmembrane domain, and a cytoplasmic Toll / interleukin-1 receptor (TIR) domain. sST2 lacks the transmembrane and cytoplasmic domains contained within ST2L and contains a unique nine amino acid (a.a.) C-terminal sequence (see, e.g., Kakkar et al. Nat. Rev. Drug Disc. 7:827-840, 2008). sST2 can function as a decoy receptor that inhibits soluble IL-33. The term also encompasses natural variants of ST2, such as splice variants (e.g., ST2V, which lacks the third immunoglobulin motif and has a unique hydrophobic tail, and ST2LV, which lacks the transmembrane domain of ST2L) or allelic variants (e.g., variants that confer protection against the risk of asthma or variants that confer a risk of asthma, as described herein). The amino acid sequence of exemplary human ST2 can be found, for example, under UniProtKB accession number Q01638. ST2 is part of the IL-33 receptor together with the coreceptor protein IL-1RAcP.Upon binding of IL-33 to the co-receptor interleukin-1 receptor accessory protein (IL-1RAcP), a 1:1:1 triple signaling complex is formed that promotes downstream signaling (see, for example, Lingel et al. Structure 17(10):1398-1410, 2009, and Liu et al. Proc. Natl. Acad. Sci. 110(37):14918-14924, 2013).

[0159] The "IL-33 axis" refers to a nucleic acid (e.g., a gene, or mRNA transcribed from a gene), or a polypeptide involved in IL-33 signal transduction. For example, the IL-33 axis can include the ligand IL-33, receptors (e.g., ST2 and / or IL-1RAcP), adapter molecules (e.g., MyD88), or proteins associated with receptor molecules and / or adapter molecules (e.g., kinases such as interleukin-1 receptor-associated kinase 1 (IRAK1) and interleukin-1 receptor-associated kinase 4 (IRAK4), or E3 ubiquitin ligases such as TNF receptor-associated factor 6 (TRAF6)).

[0160] "IL-33 axis-binding antagonist" refers to a molecule that inhibits the interaction between an IL-33 axis-binding partner and one or more of its binding partners. As used herein, IL-33 axis-binding antagonists include IL-33 binding antagonists, ST2 binding antagonists, and IL1RAcP binding antagonists.Exemplary IL-33 axis-binding antagonists include anti-IL-33 antibodies and antigen-binding fragments thereof (e.g., anti-IL-33 antibodies such as ANB-020 (AnaptysBio, Inc.), or any of the antibodies described in EP1725261, US8187596, WO2011 / 031600, WO2014 / 164959, WO2015 / 099175, or WO2015 / 106080, each of which is incorporated herein by reference in its entirety); polypeptides that bind to IL-33 and / or its receptor (ST2 and / or IL-1RAcP) and block ligand-receptor interaction (e.g., ST2-Fc proteins such as those described in WO2014 / 152195, which is incorporated herein by reference in its entirety; immunoadhesins, peptibodies, and soluble ST2, or derivatives thereof); anti-IL-33 receptor antibodies (e.g., anti-ST2 antibodies such as AMG-282 (Amgen) or STLM15 (Janssen), or any of the anti-ST2 antibodies described in WO2013 / 173761 and WO2013 / 165894, each of which is incorporated herein by reference in its entirety; or ST2-Fc proteins such as those described in WO2013 / 173761, WO2013 / 165894, or WO2014 / 152195, each of which is incorporated herein by reference in its entirety); and IL-33 receptor antagonists such as small molecule inhibitors, aptamers that bind to IL-33, and nucleic acids that hybridize to nucleic acid sequences of the IL-33 axis under stringent conditions (e.g., short interfering RNAs (siRNAs) or clustered regularly interspaced short palindromic repeat RNAs (CRISPR-RNAs or crRNAs), including single-stranded guide RNAs (sgRNAs) having the crRNA and tracrRNA sequences described in Mali et al. (Science. 339:823-26, 2013), which is incorporated herein by reference in its entirety).

[0161] The terms "anti-IL-33 antibody", "antibody that binds to IL-33", and "antibody that specifically binds to IL-33" refer to antibodies that can bind to IL-33 with sufficient affinity such that the antibody is useful for targeting IL-33 as a diagnostic and / or therapeutic agent. In one embodiment, the degree of binding of the anti-IL-33 antibody to an irrelevant non-IL-33 protein is less than about 10% of the binding of this antibody to IL-33, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the antibody that binds to IL-33 has a dissociation constant (K D ) of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g., 10 -8 M to 10 -13 M, e.g., 10 -9 M to 10 -13 M). In certain embodiments, the anti-IL-33 antibody binds to an epitope of IL-33 that is conserved among IL-33s from different species.

[0162] The term "ST2 binding antagonist" refers to a molecule that inhibits the interaction of ST2 with IL-33, IL1RAcP, and / or a second ST2 molecule. An ST2 binding antagonist can be a protein such as an "ST2-Fc protein" that includes an IL-33 binding domain (e.g., all or part of the ST2 or IL1RAcP protein) and a multimerization domain (e.g., the Fc portion of an immunoglobulin, such as isotypes IgG1, IgG2, IgG3, and IgG4, and the Fc domain of an IgG selected from any allotype within each isotype group), which are attached to each other directly or indirectly through a linker (e.g., a serine-glycine (SG) linker, a glycine-glycine (GG) linker, or variants thereof (e.g., SGG, GGS, SGS, or GSG linkers)), including the ST2-Fc proteins and their variants described in WO2013 / 173761, WO2013 / 165894, and WO2014 / 152195, which are each incorporated herein by reference in their entirety, but not limited thereto. In some embodiments, the ST2 binding antagonist can be an anti-ST2 antibody, such as AMG-282 (Amgen) or STLM15 (Janssen) or any of the anti-ST2 antibodies described in WO2013 / 173761 and WO2013 / 165894.

[0163] "Isolated nucleic acid" refers to a nucleic acid molecule separated from the components of its natural environment. An isolated nucleic acid typically includes nucleic acid molecules contained within a cell that normally contains the nucleic acid molecule, but where the nucleic acid molecule is present extrachromosomally or at a chromosomal location different from its natural chromosomal location.

[0164] The term "control sequence" refers to a DNA sequence necessary for the expression of an operably linked coding sequence in a particular host organism. For example, control sequences suitable for prokaryotes include a promoter, optionally an operator sequence, and a ribosome binding site. Eukaryotic cells are known to utilize a promoter, a polyadenylation signal, and an enhancer.

[0165] The terms "host cell", "host cell line", and "host cell culture" are used interchangeably and refer to a cell into which an exogenous nucleic acid has been introduced, including the progeny of such a cell. Host cells include "transformants" and "transformed cells", which include the primary transformed cells and progeny derived therefrom regardless of the number of passages. Progeny may contain mutations, although the nucleic acid content may not be identical to that of the parental cell. Mutant progeny having the same function or biological activity as that screened or selected for the originally transformed cell are included herein.

[0166] A nucleic acid is "operably linked" when placed into a functional relationship with another nucleic acid sequence. For example, DNA for a presequence or a secretory leader is operably linked to DNA for a polypeptide if expressed as a preprotein that participates in the secretion of the polypeptide, a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence, or a ribosome binding site is operably linked to a coding sequence if positioned to promote translation. Generally, "operably linked" means that the DNA sequences being linked are contiguous and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking can be accomplished by ligation at convenient restriction sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers are used in accordance with conventional practice.

[0167] The "percent amino acid sequence identity (%)" with respect to a polypeptide sequence identified herein is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the polypeptide being compared, after aligning the sequences to obtain the maximum percent sequence identity and introducing gaps as necessary, and not considering any conservative substitutions as part of the sequence identity. Alignments for the purpose of determining percent amino acid sequence identity can be achieved by various means within the skill in the art, such as using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. One of ordinary skill in the art can determine appropriate parameters for measuring the alignment, including any algorithms necessary to achieve the maximum alignment over the full length of the sequences being compared. However, for the purposes herein, the percent amino acid sequence identity values are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was described by Genentech, Inc., and the source code was submitted to the U.S. Copyright Office (Washington D.C., 20559) together with user documentation and is registered under U.S. Copyright No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California. The ALIGN-2 program should be compiled when used with the UNIX operating system, preferably Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.

[0168] In the context where ALIGN-2 is used for amino acid sequence comparison, the percent amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (or can be expressed as a given amino acid sequence A having or including a particular percent amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows: 100 × fraction X / Y In the formula, X is the number of amino acid residues scored as a perfect match in the alignment of A and B of the program by the array alignment program ALIGN-2, and Y is the total number of amino acid residues in B. When the length of amino acid sequence A is not equal to the length of amino acid sequence B, it is understood that the amino acid sequence identity % of A to B is not equal to the amino acid sequence identity % of B to A. Unless specifically indicated otherwise, all amino acid sequence identity % values used in this specification are obtained using the ALIGN-2 computer program as described in the immediately preceding paragraph.

[0169] The amino acid sequences described in this specification are continuous amino acid sequences unless otherwise indicated.

[0170] As used herein, the term "vector" is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid molecule to which a nucleic acid molecule is bound. One type of vector is a "plasmid", which refers to a circular double-stranded DNA loop to which additional DNA segments can be ligated. Another type of vector is a phage vector. Another type of vector is a viral vector, in which additional DNA segments are ligated into the viral genome. Certain vectors are capable of self-replication in the host cells into which they are introduced (e.g., bacterial vectors having replication of bacterial origin and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can integrate into the genome of the host cell upon introduction into the host cell and are thus replicated with the host genome. Further, certain vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "recombinant vectors" or "expression vectors"). Generally, expression vectors utilized in recombinant DNA technology are often in the form of plasmids. In this specification, "plasmid" and "vector" may be used interchangeably.

[0171] II. Compositions and Methods In one aspect, the invention is based in part on novel antibodies that bind to tryptase. In another aspect, the invention is based in part on the discovery that certain residues of anti-tryptase antibodies (e.g., HVR residues such as HVR-H3 W100 of the VH domain of the anti-tryptase antibody hu31A.v11) can be oxidation-sensitive. The invention provides pharmaceutical compositions comprising antioxidants (e.g., N-acetyltryptophan and / or methionine) to reduce or prevent oxidation of the antibodies described herein (e.g., anti-tryptase antibodies). Other suitable antioxidant excipients include, without limitation, free tryptophan, cyclodextrin, trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), pyridoxine, polyols (e.g., mannitol), and metal chelating agents (e.g., EDTA). See, for example, Ji et al., Biotechnology 98:4485-4500, 2009. The antibodies and pharmaceutical compositions of the invention are useful for the diagnosis and / or treatment of disorders (e.g., lung disorders, autoimmune deficiencies, inflammatory disorders, fibrotic disorders, granulocytic (neutrophilic or eosinophilic) disorders, monocytic disorders, lymphocytic disorders, or disorders associated with increased numbers or distribution of normal or abnormal tissue resident cells (such as mast cells, macrophages, or lymphocytes) or stromal cells (such as fibroblasts, myofibroblasts, smooth muscle cells, epithelium, or endothelium), or tryptase-related disorders or tryptase-mediated disorders. In another aspect, the invention provides a lyophilized pharmaceutical composition for reducing or eliminating oxidation of the antibodies described herein (e.g., anti-tryptase antibodies).

[0172] A. Exemplary Anti-Tryptase Antibodies The invention provides isolated antibodies that bind to tryptase. In certain embodiments, the anti-tryptase antibodies of the invention have a K of about 100 nM or less (e.g., 100 nM or less, 10 nM or less, 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less). Dand binds to trypsin. In some embodiments, the antibody has a K of 10 nM or less (e.g., 10 nM or less, 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less) D and binds to trypsin. In some embodiments, the antibody has a K of 1 nM or less (e.g., 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less) D and binds to trypsin. In some embodiments, the antibody has a K of 0.5 nM or less (e.g., 0.5 nM or less, 400 pM or less, 300 pM or less, 200 pM or less, 100 pM or less, 50 pM or less, 25 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less) D and binds to trypsin. In some embodiments, the antibody has a K of about 0.1 nM to about 0.5 nM (e.g., about 0.1 nM, about 0.2 nM, about 0.3 nM, about 0.4 nM, or about 0.5 nM) DIt binds to trypsin. In some embodiments, the antibody has a K of about 1 pM to about 500 pM, about 1 pM to about 400 pM, about 1 pM to about 300 pM, about 1 pM to about 200 pM, about 1 pM to about 100 pM, about 1 pM to about 50 pM, about 25 pM to about 500 pM, about 25 pM to about 400 pM, about 25 pM to about 300 pM, about 25 pM to about 100 pM, about 50 pM to about 500 pM, about 50 pM to about 450 pM, about 50 pM to about 425 pM, about 50 pM to about 400 pM, about 50 pM to about 375 pM, about 50 pM to about 350 pM, about 50 pM to about 325 pM, about 50 pM to about 300 pM, about 50 pM to about 275 pM, about 50 pM to about 250 pM, about 50 pM to about 200 pM, about 50 pM to about 180 pM, about 50 pM to about 175 pM, about 50 pM to about 150 pM, about 50 pM to about 125 pM, about 50 pM to about 75 pM, about 100 pM to about 500 pM, about 100 pM to about 475 pM, about 100 pM to about 450 pM, about 100 pM to about 425 pM, about 100 pM to about 400 pM, about 100 pM to about 375 pM, about 100 pM to about 350 pM, about 100 pM to about 325 pM, about 100 pM to about 300 pM, about 100 pM to about 275 pM, about 100 pM to about 250 pM, about 100 pM to about 225 pM, about 100 pM to about 200 pM, about 100 pM to about 180 pM, about 100 pM to about 175 pM, about 100 pM to about 150 pM, about 100 pM to about 125 pM, about 150 pM to about 500 pM, about 150 pM to about 475 pM, about 150 pM to about 450 pM, about 150 pM to about 425 pM, about 150 pM to about 400 pM, about 150 pM to about 375 pM, about 150 pM to about 350 pM, about 150 pM to about 325 pM, about 150 pM to about 300 pM, about 150 pM to about 375 pM, about 150 pM to about 350 pM, about 150 pM to about 325 pM, about 150 pM to about 300 pM, about 150 pM to about 275 pM, about 150 pM to about 225 pM, about 150 pM to about 200 pM, about 175 pM to about 500 pM, about 175 pM to about 475 pM, about 175 pM to about 450 pM, about 175 pM to about 425 pM, about 175 pM to about 400 pM, about 175 pM to about 375 pM, about 175 pM to about 350 pM, about 175 pM to about 325 pM, about 175 pM to about 300 pM, or about 180 pM to about 400 pM DBinds to trypsin. In some embodiments, the antibody has a K of about 0.4 nM D Binds to trypsin. In some embodiments, the antibody has a K of about 0.2 nM D Binds to trypsin. In some embodiments, the antibody has a K of about 0.18 nM D Binds to trypsin. In some embodiments, the trypsin is human trypsin, such as human trypsin beta (e.g., human trypsin beta 1, human trypsin beta 2, and / or human trypsin beta 3). In some embodiments, K D Is determined by a BIACORE® SPR assay. In certain embodiments, the trypsin is human trypsin alpha. In certain embodiments, the antibody is a human or humanized antibody.

[0173] In another embodiment, in some embodiments, the anti - tryptase antibody of the present invention (including any of the aforementioned anti - tryptase antibodies) can inhibit the activity of tryptase. In some embodiments, the anti - tryptase antibody of the present invention can inhibit the proteolytic activity of tryptase, as determined, for example, in an in vitro tryptase enzyme assay. In some embodiments, an artificial substrate, such as the synthetic peptide S - 2288, can be used as a substrate in an in vitro tryptase enzyme assay. In some embodiments, the anti - tryptase antibody of the present invention can inhibit the activity of human tryptase with a half - inhibitory concentration (IC50) of about 100 nM or less (e.g., 100 nM or less, 10 nM or less, 5 nM or less, 2.5 nM or less, 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less) when determined by, for example, an in vitro tryptase enzyme assay using S - 2288 as a substrate. In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 10 nM or less (e.g., 10 nM or less, 5 nM or less, 2.5 nM or less, 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less) when determined by, for example, an in vitro tryptase enzyme assay using S - 2288 as a substrate. In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 2.5 nM or less (e.g., 2.5 nM or less, 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less) when determined by, for example, an in vitro tryptase enzyme assay using S - 2288 as a substrate. In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 0.1 nM to about 2 nM (e.g., about 0.1 nM, about 0.2 nM, about 0.3 nM, about 0.4 nM, about 0.5 nM, about 0.6 nM, about 0.7 nM, about 0.8 nM, about 0.9 nM, about 1.0 nM, about 1.1 nM, about 1.2 nM, about 1.3 nM, about 1.4 nM, about 1.5 nM, about 1.6 nM, about 1.7 nM, about 1.8 nM, about 1.9 nM, or about 2.0 nM).In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of from about 0.5 nM to about 2.5 nM (e.g., about 0.5 nM, about 0.6 nM, about 0.7 nM, about 0.8 nM, about 0.9 nM, about 1.0 nM, about 1.1 nM, about 1.2 nM, about 1.3 nM, about 1.4 nM, about 1.5 nM, about 1.6 nM, about 1.7 nM, about 1.8 nM, about 1.9 nM, about 2.0 nM, about 2.1 nM, about 2.2 nM, about 2.3 nM, about 2.4 nM, or about 2.5 nM).In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 1 pM to about 2.5 nM, about 25 pM to about 2.5 nM, about 50 pM to about 2.5 nM, about 75 pM to about 2.5 nM, about 100 pM to about 2.5 nM, about 125 pM to about 2.5 nM, about 150 pM to about 2.5 nM, about 175 pM to about 2.5 nM, about 200 pM to about 2.5 nM, about 225 pM to about 2.5 nM, about 250 pM to about 2.5 nM, about 300 pM to about 2.5 nM, about 325 pM to about 2.5 nM, about 350 pM to about 2.5 nM, about 375 pM to about 2.5 nM, about 400 pM to about 2.5 nM, about 425 pM to about 2.5 nM, about 450 pM to about 2.5 nM, about 500 pM to about 2.5 nM, about 550 pM to about 2.5 nM, about 600 pM to about 2.5 nM, about 650 pM to about 2.5 nM, about 700 pM to about 2.5 nM, about 750 pM to about 2.5 nM, about 800 pM to about 2.5 nM, about 850 pM to about 2.5 nM, about 900 pM to about 2.5 nM, about 950 pM to about 2.5 nM, about 1 nM to about 2.5 nM, about 1.1 nM to about 2.5 nM, about 1.2 nM to about 2.5 nM, about 1.3 nM to about 2.5 nM, about 1.4 nM to about 2.5 nM, about 1.5 nM to about 2.5 nM, about 1.6 nM to about 2.5 nM, about 1.7 nM to about 2.5 nM, about 1.8 nM to about 2.5 nM, about 1.9 nM to about 2.5 nM, about 2.0 nM to about 2.5 nM, about 2.1 nM to about 2.5 nM, about 2.2 nM to about 2.5 nM, about 2.3 nM to about 2.5 nM, about 500 pM to about 1.9 pM, about 750 pM to about 1.9 pM, about 1 nM to about 1.9 pM, about 1.25 nM to about 1.9 pM, about 1.5 nM to about 1.9 pM, about 1 nM to about 1.85 nM, about 1.25 nM to about 1.85 nM, about 1.5 nM to about 1.85 nM, about 1 nM to about 1.8 nM, about 1.25 nM to about 1.8 nM, about 1.5 nM to about 1.8 nM, or about 1.6 nM to about 1.8 nM. In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 1.8 nM.In other embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of from about 0.5 nM to about 1 nM (e.g., about 0.5 nM, about 0.6 nM, about 0.7 nM, about 0.8 nM, about 0.9 nM, or about 1.0 nM).In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 1 pM to about 1 nM, about 25 pM to about 1 nM, about 50 pM to about 1 nM, about 75 pM to about 1 nM, about 100 pM to about 1 nM, about 125 pM to about 1 nM, about 150 pM to about 1 nM, about 175 pM to about 1 nM, about 200 pM to about 1 nM, about 225 pM to about 1 nM, about 250 pM to about 1 nM, about 300 pM to about 1 nM, about 325 pM to about 1 nM, about 350 pM to about 1 nM, about 375 pM to about 1 nM, about 400 pM to about 1 nM, about 425 pM to about 1 nM, about 450 pM to about 1 nM, about 500 pM to about 1 nM, about 450 pM to about 1 nM, about 500 pM to about 1 nM, about 550 pM to about 1 nM, about 600 pM to about 1 nM, about 650 pM to about 1 nM, about 700 pM to about 1 nM, about 750 pM, about 250 pM to about 800 pM, about 300 pM to about 800 pM, about 325 pM to about 800 pM, about 325 pM to about 800 pM, about 350 pM to about 800 pM, about 375 pM to about 800 pM, about 400 pM to about 800 pM, about 425 pM to about 800 pM, about 450 pM to about 800 pM, about 500 pM to about 800 pM, about 450 pM to about 800 pM, about 500 pM to about 800 pM, about 550 pM to about 800 pM, about 600 pM to about 800 pM, about 650 pM to about 800 pM, about 700 pM to about 800 pM, about 750 pM to about 800 pM, about 1 pM to about 600 pM, about 25 pM to about 600 pM, about 50 pM to about 600 pM, about 75 pM to about 600 pM, about 100 pM to about 600 pM, about 125 pM to about 600 pM, about 150 pM to about 600 pM, about 175 pM to about 600 pM, about 200 pM to about 600 pM, about 225 pM to about 600 pM, about 250 pM to about 600 pM, about 300 pM to about 600 pM, about 325 pM to about 600 pM, about 325 pM to about 600 pM, about 350 pM to about 600 pM, about 375 pM to about 600 pM, about 400 pM to about 600 pM, about 425 pM to about 600 pM, about 450 pM to about 600 pM, about 500 pM to about 600 pM, about 450 pM to about 600 pM, about 500 pM to about 600 pM, or about 550 pM to about 600 pM. In some embodiments, the antibody can inhibit the activity of human tryptase with an IC50 of about 0.6 nM.In some embodiments, the tryptase is human tryptase, such as human tryptase beta (e.g., human tryptase beta 1, human tryptase beta 2, and / or human tryptase beta 3). In some examples, the inhibitory activity of the antibody is determined as described herein, for example, by the examples (e.g., Example 1, specifically section (A)(viii)(a)), or other approaches known in the art. In certain embodiments, the antibody is a human or humanized antibody. In some embodiments, the antibody can inhibit the activity of human tryptase as a monovalent antibody or an antigen-binding antibody fragment thereof (e.g., Fab). In other embodiments, the antibody can inhibit the activity of human tryptase as a bivalent antibody (e.g., an IgG antibody (e.g., an IgG1 or IgG4 antibody) or F(ab’)2).

[0174] In some examples, any of the anti-tryptase antibodies described herein can inhibit tryptase-stimulated contraction of human primary airway smooth muscle cells. In other examples, any of the anti-tryptase antibodies described herein can inhibit tryptase-stimulated contraction of human primary airway smooth muscle cells. In still other examples, any of the anti-tryptase antibodies described herein can inhibit tryptase- or IgE-stimulated mast cell degranulation and / or histamine release. In further examples, any of the anti-tryptase antibodies described herein can, for example, reduce the amount of active tryptase (e.g., in a sample such as a bronchoalveolar lavage fluid or a nasal absorption sample) upon administration to a subject. For example, any of the anti-tryptase antibodies described herein can reduce the amount of active tryptase by about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 75%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or more. The reduction can be related to a reference amount of active tryptase, e.g., the amount of active tryptase in a sample before administration of the anti-tryptase antibody.

[0175] In some examples, the antibody (e.g., an anti-tryptase antibody) comprises at least one, two, three, four, five, or six hypervariable regions (HVRs) selected from: (a) HVR-H1 comprising the amino acid sequence of X1X2GMX3 (SEQ ID NO: 1) where X1 is Asp or Ser, X2 is Tyr or Phe, and X3 is Val or His; (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of RX1X2X3DWYFDV (SEQ ID NO: 3) where X1 is Asn or Asp, X2 is Tyr or Asn, and X3 is Asp or Tyr; (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6), or one or more combinations of the foregoing HVRs, and one or more variants thereof having at least about 80% sequence identity (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any of SEQ ID NOs: 1-6.

[0176] For example, in some embodiments, the antibody (e.g., an anti-tryptase antibody) comprises at least 1, 2, 3, 4, 5, or 6 hypervariable regions (HVRs) selected from: (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7); (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8); (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6), or one or more combinations of the foregoing HVRs, and one or more variants thereof having at least about 80% sequence identity (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 2 or 4-8.

[0177] In one particular embodiment, in some embodiments, the antibody (e.g., an anti-tryptase antibody) can include (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8), (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some embodiments, the antibody (e.g., an anti-tryptase antibody) includes (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having that sequence, (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90% identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having that sequence, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody (e.g., an anti-tryptase antibody) includes one, two, three, or four of the following heavy chain framework regions (FR): (a) FR-H1 comprising the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFS (SEQ ID NO: 11), (b) FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 12), (c) FR-H3 comprising the amino acid sequence of RFTISRDNSKNTLYLQMNSLRAEDTAVYYCTR (SEQ ID NO: 13), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14).In some embodiments, the antibody (e.g., an anti - tryptase antibody) comprises one, two, three, or four of the following light - chain FRs: (a) FR - L1 comprising the amino - acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 15), (b) FR - L2 comprising the amino - acid sequence of WYQQKPGKSPKPWIY (SEQ ID NO: 16), (c) FR - L3 comprising the amino - acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 17), and (d) FR - L4 comprising the amino - acid sequence of FGQGTKVEIK (SEQ ID NO: 18). In some embodiments, the antibody (e.g., an anti - tryptase antibody) comprises a VH domain comprising the amino - acid sequence of SEQ ID NO: 9, such as the sequence of antibody hu31A.v11, and a VL domain comprising the amino - acid sequence of SEQ ID NO: 10.

[0178] In another specific embodiment, in some embodiments, the antibody (e.g., an anti-tryptase antibody) may comprise: (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7); (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of RDNYDWYFDV (SEQ ID NO: 29); (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some embodiments, the antibody (e.g., an anti-tryptase antibody) may comprise: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of SEQ ID NO: 19, or an amino acid sequence having such sequence; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90% identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of SEQ ID NO: 20, or an amino acid sequence having such sequence; or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the antibody (e.g., an anti-tryptase antibody) comprises one, two, three, or four of the following heavy chain framework regions (FR): (a) FR-H1 comprising the amino acid sequence of EVKLVESGGGSVQPGGSRKLSCAASGFTFS (SEQ ID NO: 21); (b) FR-H2 comprising the amino acid sequence of WVRQAPGKGLEWVA (SEQ ID NO: 22); (c) FR-H3 comprising the amino acid sequence of RFTISRDNPKNTLFLQMSSLRSEDTAMYYCAR (SEQ ID NO: 23); and (d) FR-H4 comprising the amino acid sequence of WGTGTTVTVSS (SEQ ID NO: 24).In some embodiments, the antibody (e.g., an anti - tryptase antibody) comprises one, two, three, or four of the following light - chain FRs: (a) FR - L1 comprising the amino - acid sequence of QIVLTQSPAIMSASPGEKVTISC (SEQ ID NO: 25), (b) FR - L2 comprising the amino - acid sequence of WYQQKPGSSPKPWIY (SEQ ID NO: 26), (c) FR - L3 comprising the amino - acid sequence of GVPARFSGSGSGTSYSLTISSMEAEDAATYYC (SEQ ID NO: 27), and (d) FR - L4 comprising the amino - acid sequence of FGAGTKLELK (SEQ ID NO: 28). In some embodiments, the antibody (e.g., an anti - tryptase antibody) comprises a VH domain comprising the amino - acid sequence of SEQ ID NO: 19, such as the sequence of antibody 31a, and a VL domain comprising the amino - acid sequence of SEQ ID NO: 20.

[0179] In some examples, an antibody (e.g., an anti-tryptase antibody) comprises (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with any one of the amino acid sequences of SEQ ID NOs: 9, 101, 102, 103, and 104, or an amino acid sequence having such a sequence, (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90% identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with any one of the amino acid sequences of SEQ ID NOs: 10, 105, and 106, or an amino acid sequence having such a sequence, or (c) a VH domain as in (a) and a VL domain as in (b). For example, in some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 98 and a VL domain comprising the amino acid sequence of SEQ ID NO: 102. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 98 and a VL domain comprising the amino acid sequence of SEQ ID NO: 10. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 98 and a VL domain comprising the amino acid sequence of SEQ ID NO: 103. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 99 and a VL domain comprising the amino acid sequence of SEQ ID NO: 102. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 99 and a VL domain comprising the amino acid sequence of SEQ ID NO: 10. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 99 and a VL domain comprising the amino acid sequence of SEQ ID NO: 103. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 100 and a VL domain comprising the amino acid sequence of SEQ ID NO: 102. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 100 and a VL domain comprising the amino acid sequence of SEQ ID NO: 10. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 100 and a VL domain comprising the amino acid sequence of SEQ ID NO: 103. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 9 and a VL domain comprising the amino acid sequence of SEQ ID NO: 102.In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 9 and a VL domain comprising the amino acid sequence of SEQ ID NO: 10. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 9 and a VL domain comprising the amino acid sequence of SEQ ID NO: 103. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 101 and a VL domain comprising the amino acid sequence of SEQ ID NO: 102. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 101 and a VL domain comprising the amino acid sequence of SEQ ID NO: 10. In other examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 101 and a VL domain comprising the amino acid sequence of SEQ ID NO: 103.

[0180] In some examples, any of the foregoing antibodies binds to an epitope on human tryptase beta 1 that comprises at least 1, at least 2, at least 3, or all 4 residues selected from the group consisting of His51, Val80, Lys81, and Asp82 of SEQ ID NO: 71. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 that comprises at least 1, at least 2, at least 3, or all 4 residues selected from the group consisting of His51, Val80, Lys81, and Asp82 of SEQ ID NO: 71. In some embodiments, the antibody binds to an epitope on human tryptase beta 1 that comprises His51 of SEQ ID NO: 71 and at least 1, at least 2, or all 3 residues selected from the group consisting of Val80, Lys81, and Asp82. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or all 12 amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises His51, Gln67, Val80, Lys81, Asp82, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71. In some embodiments, the epitope is associated with the human tryptase beta 1 monomer or tetramer. In some embodiments, the epitope is determined by an X-ray crystallography model.In some embodiments, the antibody can dissociate both the small interface of tetrameric human tryptase beta 1 and the large interface of tetrameric human tryptase beta 1.

[0181] In some examples, any of the aforementioned anti-tryptase antibodies comprises a paratope that binds to a tryptase (e.g., human tryptase beta 1) comprising one or more amino acid residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 amino acid residues) selected from the group consisting of light chain variable region amino acid residues Val30, Thr31, Tyr32, Tyr34, Arg50, Tyr90, His92, Ser93, and Tyr94, and heavy chain variable region amino acid residues Phe50, Ser52, Gly53, Ser54, Ser55, Thr56, Tyr58, Arg95, Tyr97, and Asp98.

[0182] For example, in some examples, the anti-tryptase antibody comprises a paratope that binds to a tryptase (e.g., human tryptase beta 1) comprising light chain variable region amino acid residues Val30, Thr31, Tyr32, Tyr34, Arg50, Tyr90, His92, Ser93, and Tyr94 or heavy chain variable region amino acid residues Phe50, Ser52, Gly53, Ser54, Ser55, Thr56, Tyr58, Arg95, Tyr97, and Asp98. In some examples, the anti-tryptase antibody comprises a paratope that binds to a tryptase (e.g., human tryptase beta 1) comprising light chain variable region amino acid residues Val30, Thr31, Tyr32, Tyr34, Arg50, Tyr90, His92, Ser93, and Tyr94, and heavy chain variable region amino acid residues Phe50, Ser52, Gly53, Ser54, Ser55, Thr56, Tyr58, Arg95, Tyr97, and Asp98.

[0183] In some examples, the antibody (e.g., an anti-tryptase antibody) comprises at least one, two, three, four, five, or six hypervariable regions (HVRs) selected from (a) HVR-H1 comprising the amino acid sequence of GYAIT (SEQ ID NO: 30), (b) HVR-H2 comprising the amino acid sequence of GISSAATTFYSSWAKS (SEQ ID NO: 31), (c) HVR-H3 comprising the amino acid sequence of DPRGYGAALDRLDL (SEQ ID NO: 32), (d) HVR-L1 comprising the amino acid sequence of QSIKSVYNNRLG (SEQ ID NO: 33), (e) HVR-L2 comprising the amino acid sequence of ETSILTS (SEQ ID NO: 34), and (f) HVR-L3 comprising the amino acid sequence of AGGFDRSGDTT (SEQ ID NO: 35), or one or more combinations of the above HVRs, and one or more variants thereof having at least about 80% sequence identity (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 30-35.

[0184] In some examples, the antibody (e.g., an anti-tryptase antibody) comprises a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of the amino acid sequences of SEQ ID NOs: 36, 47, 48, 49, 50, 51, and 52, or an amino acid sequence having such a sequence, a light chain variable (VL) domain comprising an amino acid sequence having at least 90% identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of the amino acid sequences of SEQ ID NOs: 37, 53, 58, or 59, or an amino acid sequence having such a sequence, or (c) a VH domain as in (a) and a VL domain as in (b).

[0185] In some examples, any of the aforementioned antibodies (e.g., an anti-tryptase antibody) may include one, two, three, or four of the following heavy chain framework regions (FRs): (a) FR-H1 comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of EVQLVESGPGLVKPSETLSLTCTVSRFSLI (SEQ ID NO: 38), or an amino acid sequence having that sequence; (b) FR-H2 comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of WX1RQPPGKGLEWIG (SEQ ID NO: 39) where X1 is Ile or Val, or an amino acid sequence having that sequence; (c) FR-H3 comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of RX1TISX2DTSKNQX3SLKLSSVTAADTAVYX4CAR (SEQ ID NO: 40) where X1 is Val or Ser, X2 is Arg or Val, X3 is Val or Phe, and X4 is Tyr or Phe, or an amino acid sequence having that sequence; and (d) FR-H4 comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41), or an amino acid sequence having that sequence.

[0186] For example, in some examples, any of the aforementioned antibodies (e.g., anti-tryptase antibodies) may include one, two, three, or four of the following heavy chain FRs: (a) FR-H1 containing the amino acid sequence of EVQLVESGPGLVKPSETLSLTCTVSRFSLI (SEQ ID NO: 38), (b) FR-H2 containing the amino acid sequence of WIRQPPGKGLEWIG (SEQ ID NO: 42), (c) FR-H3 containing the amino acid sequence of RVTISRDTSKNQVSLKLSSVTAADTAVYYCAR (SEQ ID NO: 43), and (d) FR-H4 containing the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41).

[0187] In another example, in some examples, any of the aforementioned antibodies (e.g., anti-tryptase antibodies) may include one, two, three, or four of the following heavy chain FRs: (a) FR-H1 containing the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAVSRFSLI (SEQ ID NO: 44), (b) FR-H2 containing the amino acid sequence of WVRQAPGKGLEWIG (SEQ ID NO: 45), (c) FR-H3 containing the amino acid sequence of RSTISRDTSKNTVYLQMNSLRAEDTAVYFCAR (SEQ ID NO: 46), and (d) FR-H4 containing the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41).

[0188] In another example, in some examples, any of the aforementioned antibodies (e.g., anti-tryptase antibodies) may include one, two, three, or four of the following heavy chain FRs: (a) FR-H1 containing the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAVSRFSLI (SEQ ID NO: 44), (b) FR-H2 containing the amino acid sequence of WVRQAPGKGLEWIG (SEQ ID NO: 45), (c) FR-H3 containing the amino acid sequence of RSTISRDTSKNTVYLQMNSLRAEDTAVYFCAR (SEQ ID NO: 46), and (d) FR-H4 containing the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41).

[0189] In some examples, any of the aforementioned antibodies (e.g., anti-tryptase antibodies) may include one, two, three, or four of the following light chain FRs: (a) FR-L1 including an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of DX1QX2TQSPSSLSASVGDRVTITC (SEQ ID NO: 60) where X1 is Ile or Ala and X2 is Met or Leu, or an amino acid sequence having that sequence; (b) FR-L2 including an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of WYQQKPGKX1PKLLIY (SEQ ID NO: 61) where X1 is Ala or Pro, or an amino acid sequence having that sequence; (c) FR-L3 including an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of VPSRFSGSGSX1TDFTLTISSLQPEDFATYX2C (SEQ ID NO: 62) where X1 is Gly or Glu and X2 is Tyr or Phe, or an amino acid sequence having that sequence; and (d) FR-L4 including an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 63), or an amino acid sequence having that sequence.

[0190] For example, in certain examples, any of the aforementioned antibodies (e.g., anti-tryptase antibodies) may include one, two, three, or four of the following light chain FRs: (a) FR-L1 including the amino acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 64); (b) FR-L2 including the amino acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 65); (c) FR-L3 including the amino acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 66); and (d) FR-L4 including the amino acid sequence of FGQGTKVEIK (SEQ ID NO: 63).

[0191] In some embodiments, any one of the aforementioned antibodies (e.g., anti-tryptase antibodies) may include one, two, three, or four of the following light chain FRs: (a) FR-L1 comprising the amino acid sequence of AAVLTQTPASVSAAVGGTVSISC (SEQ ID NO: 67), (b) FR-L2 comprising the amino acid sequence of WYQQKPGQPPKLLIY (SEQ ID NO: 68), (c) FR-L3 comprising the amino acid sequence of GVPSRFKGSGSETQFTLTISDVQX1DDAATYFC (SEQ ID NO: 69) wherein X1 is Cys or Ala, and (d) FR-L4 comprising the amino acid sequence of FGQGTKVEIKFGGGTEVVVK (SEQ ID NO: 70).

[0192] In some examples, an antibody (e.g., an anti-tryptase antibody) comprises (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of the amino acid sequences of SEQ ID NOs: 36, 47, 48, 49, 50, 51, and 52, or an amino acid sequence having such a sequence, (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90% identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of the amino acid sequences of SEQ ID NOs: 37, 53, 58, or 59, or an amino acid sequence having such a sequence, or (c) a VH domain as in (a) and a VL domain as in (b). For example, in some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 36 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 47 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 48 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 49 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 50 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 50 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 51 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 52 and a VL domain comprising the amino acid sequence of SEQ ID NO: 37. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 36 and a VL domain comprising the amino acid sequence of SEQ ID NO: 53.In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 47 and a VL domain comprising the amino acid sequence of SEQ ID NO: 53. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 48 and a VL domain comprising the amino acid sequence of SEQ ID NO: 53. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 49 and a VL domain comprising the amino acid sequence of SEQ ID NO: 53. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 50 and a VL domain comprising the amino acid sequence of SEQ ID NO: 53. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 51 and a VL domain comprising the amino acid sequence of SEQ ID NO: 53. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 52 and a VL domain comprising the amino acid sequence of SEQ ID NO: 53. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 36 and a VL domain comprising the amino acid sequence of SEQ ID NO: 58. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 47 and a VL domain comprising the amino acid sequence of SEQ ID NO: 58. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 48 and a VL domain comprising the amino acid sequence of SEQ ID NO: 58. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 49 and a VL domain comprising the amino acid sequence of SEQ ID NO: 58. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 50 and a VL domain comprising the amino acid sequence of SEQ ID NO: 58. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 51 and a VL domain comprising the amino acid sequence of SEQ ID NO: 58. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 36 and a VL domain comprising the amino acid sequence of SEQ ID NO: 59. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 47 and a VL domain comprising the amino acid sequence of SEQ ID NO: 59. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 48 and a VL domain comprising the amino acid sequence of SEQ ID NO: 59.In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 49 and a VL domain comprising the amino acid sequence of SEQ ID NO: 59. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 50 and a VL domain comprising the amino acid sequence of SEQ ID NO: 59. In some examples, the antibody comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 51 and a VL domain comprising the amino acid sequence of SEQ ID NO: 59.

[0193] In another specific embodiment, in some embodiments, the anti-tryptase antibody may comprise: (a) HVR-H1 comprising the amino acid sequence of GYAIT (SEQ ID NO: 30), (b) HVR-H2 comprising the amino acid sequence of GISSAATTFYSSWAKS (SEQ ID NO: 31), (c) HVR-H3 comprising the amino acid sequence of DPRGYGAALDRLDL (SEQ ID NO: 32), (d) HVR-L1 comprising the amino acid sequence of QSIKSVYNNRLG (SEQ ID NO: 33), (e) HVR-L2 comprising the amino acid sequence of ETSILTS (SEQ ID NO: 34), and (f) HVR-L3 comprising the amino acid sequence of AGGFDRSGDTT (SEQ ID NO: 35). In some embodiments, the anti-tryptase antibody comprises: (a) a heavy chain variable (VH) domain comprising the amino acid sequence of SEQ ID NO: 36 and at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) thereto, or an amino acid sequence having such a sequence, (b) a light chain variable (VL) domain comprising the amino acid sequence of SEQ ID NO: 37 and at least 90% identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) thereto, or an amino acid sequence having such a sequence, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, the anti-tryptase antibody comprises one, two, three, or four of the following heavy chain framework regions (FR): (a) FR-H1 comprising the amino acid sequence of EVQLVESGPGLVKPSETLSLTCTVSRFSLI (SEQ ID NO: 38), (b) FR-H2 comprising the amino acid sequence of WIRQPPGKGLEWIG (SEQ ID NO: 42), (c) FR-H3 comprising the amino acid sequence of RVTISRDTSKNQVSLKLSSVTAADTAVYYCAR (SEQ ID NO: 43), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 41).In some embodiments, the anti - tryptase antibody comprises one, two, three, or four of the following light - chain FRs: (a) FR - L1 comprising the amino - acid sequence of DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 64), (b) FR - L2 comprising the amino - acid sequence of WYQQKPGKAPKLLIY (SEQ ID NO: 65), (c) FR - L3 comprising the amino - acid sequence of GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 66), and (d) FR - L4 comprising the amino - acid sequence of FGQGTKVEIK (SEQ ID NO: 63). In some embodiments, the anti - tryptase antibody comprises a VH domain comprising the amino - acid sequence of SEQ ID NO: 36, such as the anti - tryptase antibody huE104.v2, and a VL domain comprising the amino - acid sequence of SEQ ID NO: 37.

[0194] In another specific embodiment, in some embodiments, an antibody (e.g., an anti-tryptase antibody) may comprise (a) HVR-H1 comprising the amino acid sequence of GYAIT (SEQ ID NO: 30), (b) HVR-H2 comprising the amino acid sequence of GISSAATTFYSSWAKS (SEQ ID NO: 31), (c) HVR-H3 comprising the amino acid sequence of DPRGYGAALDRLDL (SEQ ID NO: 32), (d) HVR-L1 comprising the amino acid sequence of QSIKSVYNNRLG (SEQ ID NO: 33), (e) HVR-L2 comprising the amino acid sequence of ETSILTS (SEQ ID NO: 34), and (f) HVR-L3 comprising the amino acid sequence of AGGFDRSGDTT (SEQ ID NO: 35). In some embodiments, an antibody (e.g., an anti-tryptase antibody) comprises (a) a heavy chain variable (VH) domain comprising the amino acid sequence of SEQ ID NO: 52 and at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity), or an amino acid sequence having that sequence, (b) a light chain variable (VL) domain comprising the amino acid sequence of SEQ ID NO: 53 and at least 90% identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity), or an amino acid sequence having that sequence, or (c) a VH domain as in (a) and a VL domain as in (b). In some embodiments, an antibody (e.g., an anti-tryptase antibody) comprises one, two, three, or four of the following heavy chain framework regions (FR): (a) FR-H1 comprising the amino acid sequence of QXSLEESGGGLFKPTDTLTLTCTVSRFSLI (SEQ ID NO: 54), (b) FR-H2 comprising the amino acid sequence of WVRQSPENGLEWIG (SEQ ID NO: 55), (c) FR-H3 comprising the amino acid sequence of RSTITRNTNENTVTLKMTSLTAADTATYFCAR (SEQ ID NO: 56), and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 57).In some embodiments, the antibody (e.g., an anti - tryptase antibody) comprises one, two, three, or four of the following light - chain FRs: (a) FR - L1 comprising the amino - acid sequence of AAVLTQTPASVSAAVGGTVSISC (SEQ ID NO: 67), (b) FR - L2 comprising the amino - acid sequence of WYQQKPGQPPKLLIY (SEQ ID NO: 68), (c) FR - L3 comprising the amino - acid sequence of GVPSRFKGSGSETQFTLTISDVQX1DDAATYFC (SEQ ID NO: 69) where X1 is Cys or Ala, and (d) FR - L4 comprising the amino - acid sequence of FGQGTKVEIKFGGGTEVVVK (SEQ ID NO: 70). In some embodiments, the antibody (e.g., an anti - tryptase antibody) comprises a VH domain comprising the amino - acid sequence of SEQ ID NO: 52, such as antibody E104, and a VL domain comprising the amino - acid sequence of SEQ ID NO: 53. In some embodiments, the antibody (e.g., an anti - tryptase antibody) comprises a VH domain comprising the amino - acid sequence of SEQ ID NO: 52 and a VL domain comprising the amino - acid sequence of SEQ ID NO: 59.

[0195] In some examples, the present invention provides an antibody comprising (a) a heavy chain comprising an amino - acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino - acid sequence of SEQ ID NO: 76, or an amino - acid sequence having that sequence, and / or (b) a light chain comprising an amino - acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino - acid sequence of SEQ ID NO: 77, or an amino - acid sequence having that sequence. In some examples, the antibody comprises a heavy chain comprising the amino - acid sequence of SEQ ID NO: 76 and a light chain comprising the amino - acid sequence of SEQ ID NO: 77. In some embodiments, the heavy chain further comprises a lysine (K) residue at the C - terminus.

[0196] In some examples, the present invention provides an antibody comprising (a) a heavy chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of SEQ ID NO: 78, or an amino sequence having such a sequence, and / or (b) a light chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of SEQ ID NO: 79, or an amino sequence having such a sequence. In some examples, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 78 and a light chain comprising the amino acid sequence of SEQ ID NO: 79. In some embodiments, the heavy chain further comprises a lysine (K) residue at the C-terminus.

[0197] In some examples, the present invention provides an antibody comprising (a) a heavy chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of SEQ ID NO: 80, or an amino sequence having such a sequence, and / or (b) a light chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with the amino acid sequence of SEQ ID NO: 81, or an amino sequence having such a sequence. In some examples, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 80 and a light chain comprising the amino acid sequence of SEQ ID NO: 81. In some embodiments, the heavy chain further comprises a lysine (K) residue at the C-terminus.

[0198] In some examples, the invention provides an antibody comprising (a) a heavy chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of SEQ ID NO: 82, or an amino sequence having such a sequence, and / or (b) a light chain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the amino acid sequence of SEQ ID NO: 83, or an amino sequence having such a sequence. In some examples, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain comprising the amino acid sequence of SEQ ID NO: 83. In some embodiments, the heavy chain further comprises a lysine (K) residue at the C-terminus.

[0199] In some examples, any one of the aforementioned antibodies binds to an epitope on human tryptase beta 1 that comprises at least 1, at least 2, or all 3 residues selected from the group consisting of Gln100, Leu101, and Leu102 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 further comprises one or more amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71. In some embodiments, the epitope on human tryptase beta 1 comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or all 14 amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71. In some embodiments, the epitope comprises Gln35, Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Gln100, Leu101, Leu102, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, Glu129, and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is related to the human tryptase beta 1 monomer or tetramer. In some embodiments, the epitope is related to the human tryptase beta 1 tetramer and the epitope on human tryptase beta 1 further comprises one or both of Gln35 and Arg216 of SEQ ID NO: 71. In some embodiments, the epitope is determined by an X-ray crystallography model. In some embodiments, the antibody can dissociate the small interface and / or the large interface of human tryptase beta 1.

[0200] In some examples, any one of the aforementioned anti-tryptase antibodies comprises a paratope that binds to a tryptase (e.g., human tryptase beta 1) comprising one or more amino acid residues selected from the group consisting of light chain variable region amino acid residues Tyr29, Asn30, Arg32, and Arg94, and heavy chain variable region amino acid residues Gly31, Tyr32, Ser52, Ser53, Ala54, Thr56, Phe58, Pro96, Arg97, Gly98, Tyr99, and Arg100e (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16, 19 amino acid residues).

[0201] For example, in some examples, the anti-tryptase antibody comprises a paratope that binds to a tryptase (e.g., human tryptase beta 1) comprising light chain variable region amino acid residues Tyr29, Asn30, Arg32, and Arg94 or heavy chain variable region amino acid residues Gly31, Tyr32, Ser52, Ser53, Ala54, Thr56, Phe58, Pro96, Arg97, Gly98, Tyr99, and Arg100e. In some examples, the anti-tryptase antibody comprises a paratope that binds to a tryptase (e.g., human tryptase beta 1) comprising light chain variable region amino acid residues Tyr29, Asn30, Arg32, and Arg94, and heavy chain variable region amino acid residues Gly31, Tyr32, Ser52, Ser53, Ala54, Thr56, Phe58, Pro96, Arg97, Gly98, Tyr99, and Arg100e.

[0202] In some examples, any one of the aforementioned anti-tryptase antibodies binds to human tryptase. In some examples, any one of the aforementioned antibodies binds to cynomolgus (cyno) tryptase. In some examples, the antibody binds to human tryptase alpha or human tryptase beta. In some examples, the antibody binds to human tryptase beta 1, human tryptase beta 2, or human tryptase beta 3.

[0203] In another aspect, the present invention provides an anti-tryptase antibody that binds to an epitope on tryptase (e.g., human tryptase beta 1) that can refer to the amino acid sequence of SEQ ID NO: 71 and that comprises one or more amino acid residues (e.g., 1, 2, 3, 4, 5, 6, or 7 amino acid residues) selected from the group consisting of His51, Val80, Lys81, Asp82, Leu83, Ala84, and Ala85, or the corresponding amino acids of any tryptase protein. For example, in some embodiments, the antibody binds to an epitope on tryptase (e.g., human tryptase beta 1) that comprises at least 1, at least 2, at least 3, or all 4 residues selected from the group consisting of His51, Val80, Lys81, and Asp82 of SEQ ID NO: 71, or the corresponding amino acids of any tryptase protein. In some embodiments, the antibody binds to an epitope on tryptase (e.g., human tryptase beta 1) that comprises His51 of SEQ ID NO: 71 and at least 1, at least 2, or all 3 residues selected from the group consisting of Val80, Lys81, and Asp82, or the corresponding amino acids of any tryptase protein. In some embodiments, the epitope on tryptase (e.g., human tryptase beta 1) further comprises one or more amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71, or the corresponding amino acids of any tryptase protein.In some embodiments, the epitope on tryptase (e.g., human tryptase beta 1) comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or all 12 amino acid residues selected from the group consisting of Gln67, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71, or the corresponding amino acids of any tryptase protein. In some embodiments, the epitope on tryptase (e.g., human tryptase beta 1) comprises His51, Gln67, Val80, Lys81, Asp82, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO: 71, or the corresponding amino acids of any tryptase protein. In some embodiments, the epitope is associated with the tryptase (e.g., human tryptase beta 1) monomer or tetramer. In some embodiments, the epitope is determined by an X-ray crystallography model. In some embodiments, the antibody is capable of dissociating both the small interface of the tetrameric tryptase and the large interface of the tetrameric tryptase tryptase (e.g., human tryptase beta 1).

[0204] In yet another aspect, the present invention provides an anti-tryptase antibody that binds to an epitope of tryptase (e.g., human tryptase beta 1) that can refer to the amino acid sequence of SEQ ID NO: 71 and includes one or more amino acid residues (e.g., 1, 2, 3, 4, 5, 6, or 7 amino acid residues) selected from the group consisting of Gln100, Leu101, Leu102, Pro103, Val104, Ser105, and Arg106, or corresponding amino acids of any tryptase protein. In some embodiments, the epitope on tryptase (e.g., human tryptase beta 1) further includes one or more amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71, or corresponding amino acids of any tryptase protein. In some embodiments, the epitope on tryptase (e.g., human tryptase beta 1) includes at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or all 14 amino acid residues selected from the group consisting of Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, and Glu129 of SEQ ID NO: 71, or corresponding amino acids of any tryptase protein. In some embodiments, the epitope includes Gln35, Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Gln100, Leu101, Leu102, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, Glu129, and Arg216 of SEQ ID NO: 71, or corresponding amino acids of any tryptase protein. In some embodiments, the epitope is associated with a tryptase (e.g., human tryptase beta 1) monomer or tetramer.In some embodiments, the epitope is associated with the tetramer, and the epitope on tryptase (e.g., human tryptase beta 1) comprises one or both of Gln35 and Arg216 of SEQ ID NO: 71, or the corresponding amino acids of any tryptase protein. In some embodiments, the epitope is determined by an X-ray crystallography model. In some embodiments, the antibody can dissociate the small and / or large interfaces of tryptase (e.g., human tryptase beta 1).

[0205] In some embodiments, any of the aforementioned antibodies binds to an epitope on tryptase (e.g., human tryptase beta 1) that comprises one or more amino acid residues (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 amino acid residues) selected from the group consisting of Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, and Glu129, which may refer to the amino acid sequence of SEQ ID NO: 71, or the corresponding amino acids of any tryptase protein.

[0206] For example, in some instances, any of the aforementioned antibodies binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Gln67 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Asp82 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Leu83 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Ala84 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Arg87 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Pro103 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Val104 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Ser105 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Arg106 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some instances, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that includes Glu128 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein.In some examples, the antibody binds to an epitope on trypsin (e.g., human trypsin beta 1) that includes Glu129 of SEQ ID NO: 71 or the corresponding amino acid of any trypsin protein.

[0207] In some embodiments, any of the foregoing antibodies may refer to the amino acid sequence of SEQ ID NO: 71 and binds to an epitope on trypsin (e.g., human trypsin beta 1) that includes two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, or all eleven amino acid residues selected from the group consisting of Gln67, Asp82, Leu83, Ala84, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, and Glu129, or the corresponding amino acids of any trypsin protein.

[0208] In some examples, any one of the aforementioned antibodies may refer to the amino acid sequence of SEQ ID NO: 71 and binds to an epitope on a tryptase that includes one or more additional amino acid residues (e.g., 1, 2, 3, 4, or 5 additional amino acid residues) selected from the group consisting of His51, Val80, Lys81, Ala85, and Pro130, or the corresponding amino acids of any tryptase protein. For example, in some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further includes His51 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further includes Val80 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some embodiments, the anti-tryptase antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further includes Lys81 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further includes Ala85 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further includes Pro130 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein.

[0209] In some examples, the antibody binds to an epitope on a tryptase that includes two or more, three or more, four or more, or five or more additional amino acid residues selected from the group consisting of His51, Val80, Lys81, Ala85, and Pro130, which may refer to the amino acid sequence of SEQ ID NO: 71, or the corresponding amino acids of any tryptase protein.

[0210] In some examples, the anti - tryptase antibody binds to an epitope on tryptase (e.g., human tryptase beta 1) that includes His51, Gln67, Val80, Lys81, Asp82, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO:71, or the corresponding amino acids of any tryptase protein. In some examples, the anti - tryptase antibody binds to an epitope on tryptase (e.g., human tryptase beta 1) consisting of His51, Gln67, Val80, Lys81, Asp82, Leu83, Ala84, Ala85, Arg87, Pro103, Val104, Ser105, Arg106, Glu128, Glu129, and Pro130 of SEQ ID NO:71.

[0211] In other examples, any of the aforementioned anti-tryptase antibodies may refer to the amino acid sequence of SEQ ID NO: 71 and bind to an epitope on a tryptase (e.g., human tryptase beta 1) that comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) additional amino acid residues selected from the group consisting of Gln35, Trp55, Gln100, Leu101, Leu102, Glu126, Leu127, and Arg216, or the corresponding amino acids of any tryptase protein. For example, in some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Gln35 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Trp55 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Gln100 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Leu101 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Leu102 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Glu126 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Leu127 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein. In some examples, the antibody binds to an epitope on a tryptase (e.g., human tryptase beta 1) that further comprises Arg216 of SEQ ID NO: 71 or the corresponding amino acid of any tryptase protein.In some examples, the antibody binds to an epitope on tryptase (e.g., human tryptase beta 1) that may refer to the amino acid sequence of SEQ ID NO: 71 and includes two or more, three or more, four or more, five or more, six or more, seven or more, or eight or more additional amino acid residues selected from the group consisting of Gln35, Trp55, Gln100, Leu101, Leu102, Glu126, Leu127, and Arg216, or the corresponding amino acids of any tryptase protein.

[0212] In some examples, the anti-tryptase antibody binds to an epitope on tryptase (e.g., human tryptase beta 1) that may refer to the amino acid sequence of SEQ ID NO: 71 and includes Gln35, Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Gln100, Leu101, Leu102, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, Glu129, and Arg216, or the corresponding amino acids of any tryptase protein. In some examples, the anti-tryptase antibody binds to an epitope on tryptase (e.g., human tryptase beta 1) consisting of Gln35, Trp55, Gln67, Asp82, Leu83, Ala84, Arg87, Gln100, Leu101, Leu102, Pro103, Val104, Ser105, Arg106, Glu126, Leu127, Glu128, Glu129, and Arg216 of SEQ ID NO: 71.

[0213] In another aspect, the present invention provides an antibody that competes with any of the aforementioned antibodies for binding to tryptase (e.g., human tryptase beta 1).

[0214] In another aspect, the present invention provides an antibody that binds to the same epitope or overlapping epitopes as any of the aforementioned antibodies.

[0215] In some embodiments, any of the aforementioned antibodies may have the ability to cleave tryptase having a tetrameric structure (such as mature tryptase present in or released from mast cell secretory granules) to form small molecular weight species, such as monomers, dimers, and / or trimers.

[0216] In a further aspect, an antibody according to any of the above embodiments may incorporate any of the features described in Sections 1-7 below, alone or in combination.

[0217] 1. Antibody affinity In certain embodiments, the antibodies provided herein have a dissociation constant (K -6 ) of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, ≦1 pM, or ≦0.1 pM (e.g., 10 -6 M or less, e.g., 10 -9 M to 10 -9 M or less, e.g., 10 -13 M or less). In some embodiments, the anti-tryptase antibodies of the invention bind to tryptase (e.g., human tryptase, e.g., human tryptase beta) with a K D of about 100 nM or less (e.g., 100 nM or less, 10 nM or less, 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less). In some embodiments, the antibody binds to tryptase (e.g., human tryptase, e.g., human tryptase beta) with a K D of 10 nM or less (e.g., 10 nM or less, 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less). In some embodiments, the antibody binds to tryptase (e.g., human tryptase, e.g., human tryptase beta) with a K D of 1 nM or less (e.g., 1 nM or less, 100 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less). In some embodiments, the antibody binds to tryptase (e.g., human tryptase, e.g., human tryptase beta) with a K DIt binds to trypsin (e.g., human trypsin, e.g., human trypsin beta). In some embodiments, any of the anti-trypsin antibodies described above or herein has a K of about 0.5 nM or less (e.g., 0.5 nM or less, 400 pM or less, 300 pM or less, 200 pM or less, 100 pM or less, 50 pM or less, 25 pM or less, 10 pM or less, 1 pM or less, or 0.1 pM or less). D It binds to trypsin (e.g., human trypsin, e.g., human trypsin beta). In some embodiments, the antibody has a K of about 0.1 nM to about 0.5 nM (e.g., about 0.1 nM, about 0.2 nM, about 0.3 nM, about 0.4 nM, or about 0.5 nM). D It binds to trypsin (e.g., human trypsin, e.g., human trypsin beta). In some embodiments, the antibody has a K of about 0.4 nM. D It binds to trypsin (e.g., human trypsin, e.g., human trypsin beta). In some embodiments, the antibody has a K of about 0.18 nM. D It binds to trypsin (e.g., human trypsin, e.g., human trypsin beta).

[0218] In one embodiment, K D is measured by a radioimmunoassay (RIA). In one embodiment, the RIA is performed using the Fab version of the antibody of interest and its antigen. For example, the solution binding affinity of the Fab for the antibody is determined in the presence of a series of titrations of unlabeled antigen at the lowest concentration of ( 125I) It is measured by equilibration with the labeled antigen followed by capture of the bound antibody on a plate coated with an anti-Fab antibody (see, for example, Chen et al. J. Mol. Biol. 293:865-881, 1999). To establish the conditions for the assay, a MICROTITER® multiwell plate (Thermo Scientific) is coated overnight with 5 μg / ml of the capture anti-Fab antibody (Cappel Labs) in 50 mM carbonate (pH 9.6), followed by blocking with 2% (w / v) bovine serum albumin in PBS for 2 - 5 hours at room temperature (about 23°C). In a non-adsorptive plate (Nunc number 269620), 100 pM or 26 pM 125 I]-antigen is mixed with serial dilutions of the Fab of interest (e.g., consistent with the evaluation of the anti-VEGF antibody, Fab-12 in Presta et al. Cancer Res. 57:4593-4599, 1997). The Fab of interest is then incubated overnight, although this incubation can be continued for a longer period (e.g., about 65 hours) to ensure that equilibrium is achieved. Thereafter, the mixture is transferred to the capture plate for incubation at room temperature (e.g., 1 hour). The solution is then removed and the plate is washed 8 times with 0.1% polysorbate 20 (TWEEN-20®) in PBS. Once the plate is dry, 150 μL / well of scintillant (MICROSCINT-20™, Packard) is added and the plate is counted for 10 minutes on a TOPCOUNT® gamma counter (Packard). The concentration of each Fab that gives a maximum binding of 20% or less is selected for use in the competitive binding assay.

[0219] According to another embodiment, K DIt is measured using a BIACORE (registered trademark) surface plasmon resonance assay. For example, an assay using a BIACORE (registered trademark)-2000 or BIACORE (registered trademark)-3000 (BIAcore, Inc., Piscataway, NJ) is performed at 25 °C using an immobilized antigen CM5 chip at about 10 response units (RU). In one embodiment, a carboxymethylated dextran biosensor chip (CM5, BIACORE, Inc.) is activated using N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. After diluting the antigen to 5 μg / ml (about 0.2 μM) with 10 mM sodium acetate at pH 4.8, it is injected at a flow rate of 5 μl / min to achieve about 10 response units (RU) of the coupled protein. Following the injection of the antigen, 1 M ethanolamine is injected to block unreacted groups. For kinetic measurements, two-fold serial dilutions of the Fab (0.78 nM to 500 nM) are injected at 25 °C at a flow rate of about 25 μL / min into phosphate buffered saline (PBS) containing 0.05% polysorbate 20 (TWEEN (registered trademark) 20) surfactant (PBST). The association rate (k on ) and the dissociation rate (k off ) are calculated by simultaneously fitting the association sensorgram and the dissociation sensorgram using a simple 1:1 Langmuir binding model (BIACORE (registered trademark) evaluation software version 3.2). The equilibrium dissociation constant (K D ) is calculated as the ratio k off / k on . See, for example, Chen et al. (J Mol. Biol. 293:865-881, 1999). The on-rate is 10 6 M -1 s -1When it exceeds, the on-rate can be determined by using a fluorescence quenching technique that measures the increase or decrease in the fluorescence emission intensity (excitation = 295 nm, emission = 340 nm, 16 nm bandpass) of 20 nM anti-antigen antibody (Fab type) in PBS (pH 7.2) at 25°C in the presence of antigen of increasing concentration, measured in a spectrometer such as a stopped-flow equipped spectrophotometer (Aviv Instruments) with a stirred cuvette or an 8000 series SLM-AMINCO™ spectrophotometer (ThermoSpectronic).

[0220] In some embodiments, K D is measured using, for example, a BIACORE® SPR assay as described in section (A)(vii) of Example 1. In some embodiments, the SPR assay can use a BIAcore® T200 or equivalent instrument. In some embodiments, a BIAcore® series S CM5 sensor chip (or equivalent sensor chip) is immobilized with a monoclonal mouse anti-human IgG (Fc) antibody, and the anti-tryptase antibody is sequentially captured on the flow cell. Serial 3-fold dilutions of His-tagged human tryptase beta 1 monomer (SEQ ID NO: 128) are injected at a flow rate of 30 μl / min. Each sample is analyzed for 3 minutes of association and 10 minutes of dissociation. The assay is performed at 25°C. After each injection, the chip is regenerated using 3 M MgCl2. The binding response is corrected by subtracting the response units (RU) from a flow cell that captures irrelevant IgG of similar density. k on and k off of the simultaneous fitting of a 1:1 Langmuir model is used for kinetic analysis.

[0221] 2. Antibody Fragments In certain embodiments, the antibodies provided herein are antibody fragments. Antibody fragments include, but are not limited to, Fab, Fab’, Fab’-SH, F(ab’)2, Fv, and scFv fragments, and other fragments described below. For an overview of certain antibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For an overview of scFv fragments, see, for example, Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenberg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994), as well as WO93 / 16185, and U.S. Patent Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab’)2 fragments that include salvage receptor binding epitope residues and have an increased in vivo half-life, see U.S. Patent No. 5,869,046.

[0222] A diabody is an antibody fragment having two antigen-binding sites that can be bivalent or bispecific. See, for example, EP404,097, WO1993 / 01161, Hudson et al. Nat. Med. 9:129-134, 2003, and Hollinger et al. Proc. Natl. Acad. Sci. USA 90:6444-6448, 1993. Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134, 2003.

[0223] A single domain antibody is an antibody fragment that includes all or a portion of the heavy chain variable domain of an antibody, or all or a portion of the light chain variable domain. In certain embodiments, the single domain antibody is a human single domain antibody (see, for example, U.S. Patent No. 6,248,516B1).

[0224] Antibody fragments can be made by a variety of techniques including, but not limited to, proteolytic cleavage of intact antibodies as described herein and production in recombinant host cells (e.g., E. coli or phage).

[0225] 3. Chimeric and Humanized Antibodies In certain embodiments, the antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567 and Morrison et al. Proc. Natl. Acad. Sci. USA, 81:6851-6855, 1984). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate such as a monkey) and a human constant region. In a further example, a chimeric antibody is a “class switch” antibody in which the class or subclass has changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0226] In certain embodiments, a chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce its immunogenicity in humans while retaining the specificity and affinity of the non-human parent antibody. Generally, a humanized antibody comprises one or more variable domains in which the HVRs (or portions thereof) are derived from a non-human antibody and the FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody optionally also comprises at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues derived from a non-human antibody (e.g., the antibody from which the HVR residues are derived) to, for example, restore or improve antibody specificity or affinity.

[0227] Humanized antibodies and methods for making them are reviewed, for example, in Almagro et al. Front. Biosci. 13:1619-1633, 2008, and are further described, for example, in Riechmann et al. Nature 332:323-329, 1988, Queen et al. Proc. Natl. Acad. Sci. USA 86:10029-10033, 1989, U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409, Kashmiri et al. Methods 36:25-34, 2005 (describing specific-determining region (SDR) grafting), Padlan, Mol. Immunol. 28:489-498, 1991 (describing "resurfacing"), Dall’Acqua et al. Methods 36:43-60, 2005 (describing "FR shuffling"), and Osbourn et al. Methods 36:61-68, 2005 and Klimka et al. Br. J. Cancer, 83:252-260, 2000 (describing an "inductive selection" approach to FR shuffling).

[0228] Human framework regions that can be used for humanization include framework regions selected using the "best fit" method (see, e.g., Sims et al. J. Immunol. 151:2296, 1993), framework regions derived from consensus sequences of human antibodies of certain subgroups of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285, 1992, and Presta et al. J. Immunol., 151:2623, 1993); human mature (somatic mutated) framework regions or human germline framework regions (see, e.g., Almagro et al. Front. Biosci. 13:1619-1633, 2008), as well as framework regions derived from screening of FR libraries (see, e.g., Baca et al. J. Biol. Chem. 272:10678-10684, 1997 and Rosok et al. J. Biol. Chem. 271:22611-22618, 1996), but are not limited thereto.

[0229] 4. Human Antibodies In certain embodiments, the antibodies provided herein are human antibodies. Human antibodies can be produced using a variety of techniques known in the art. Human antibodies are generally described in van Dijk et al. Curr. Opin. Pharmacol. 5:368-74, 2001 and Lonberg, Curr. Opin. Immunol. 20:450-459, 2008.

[0230] Human antibodies can be prepared by administering an immunogen to a transgenic animal modified to produce intact human antibodies or intact antibodies having human variable regions in response to antigen administration. Such animals typically contain all or part of a human immunoglobulin locus that replaces the endogenous immunoglobulin locus or is present extrachromosomally or randomly integrated into the chromosomes of the animal. In such transgenic mice, the endogenous immunoglobulin locus is generally inactivated. For an overview of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125, 2005. See also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584 (describing the XENOMOUSE™ technology), U.S. Patent No. 5,770,429 (describing the HUMAB® technology), U.S. Patent No. 7,041,870 (describing the K-M MOUSE® technology), and U.S. Patent Application Publication No. 2007 / 0061900 (describing the VELOCIMOUSE® technology). Human variable regions derived from intact antibodies produced by such animals can be further modified, for example, by combining with different human constant regions.

[0231] Human antibodies can also be produced by methods based on hybridomas. Human myelomas and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described. (See, e.g., Kozbor J. Immunol. 133:3001, 1984; Brodeur et al. Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al. J. Immunol. 147:86, 1991). Human antibodies generated via human B-cell hybridoma technology are also described in Li et al. Proc. Natl. Acad. Sci. USA, 103:3557-3562, 2006. Further methods include, for example, those described in U.S. Patent No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (triooma technology) is also described in Vollmers et al. Histology and Histopathology 20(3):927-937, 2005 and Vollmers et al. Methods and Findings in Experimental and Clinical Pharmacology 27(3):185-91, 2005.

[0232] Human antibodies can also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. Such variable domain sequences can then be combined with desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below.

[0233] 5. Library-Derived Antibodies The antibodies of the present invention can be isolated by screening a combinatorial library for antibodies having the desired activity(ies). For example, methods for generating phage display libraries and screening such libraries for antibodies possessing the desired binding properties are known in the art. Such methods are reviewed, for example, in Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O’Brien et al., ed., Human Press, Totowa, NJ, 2001), and are further described, for example, in McCafferty et al. Nature 348:552-554, 1990, Clackson et al. Nature 352:624-628, 1991, Marks et al. J. Mol. Biol. 222:581-597, 1992, Marks et al. in Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, NJ, 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.

[0234] In certain phage display methods, repertoires of VH and VL genes are cloned separately by polymerase chain reaction (PCR), randomly recombined in a phage library, and this can then be screened for antigen-binding phage as described in Winter et al. Ann. Rev. Immunol., 12:433-455, 1994. Phage typically display antibody fragments as either single-chain Fv (scFv) fragments or Fab fragments. Libraries derived from immunizing sources provide high-affinity antibodies to the immunogen without the need to construct hybridomas. Alternatively, as described in Griffiths et al. EMBO J. 12:725-734, 1993, naive repertoires can be cloned (e.g., from humans) to provide a single source of antibodies to a wide range of non-self and self antigens without immunization. Finally, as described in Hoogenboom et al. J. Mol. Biol., 227:381-388, 1992, unrearranged V gene segments from stem cells can be cloned and highly variable HVR3 regions encoded using PCR primers containing random sequences, and a naive library can be synthetically generated by achieving rearrangement in vitro. Patent publications describing human antibody phage libraries include, for example, U.S. Patent No. 5,750,373, as well as U.S. Patent Application Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0235] Antibodies or antibody fragments isolated from a human antibody library are considered herein to be human antibodies or human antibody fragments.

[0236] 6. Multispecific Antibodies In certain embodiments, the antibodies provided herein are multispecific antibodies, e.g., bispecific antibodies. Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different sites. In certain embodiments, the bispecific antibody can bind to two different epitopes of tryptase. In certain embodiments, one of the binding specificities is for tryptase and the other is for any other antigen (e.g., a second biological molecule). In some embodiments, the bispecific antibody can bind to two different epitopes of tryptase. In other embodiments, one of the binding specificities is for tryptase (e.g., human tryptase, e.g., human tryptase beta) and the other is for any other antigen (e.g., a second biological molecule, e.g., IL-13, IL-4, IL-5, IL-17, IL-33, IgE, M1 prime, CRTH2, or TRPA). Thus, the bispecific antibody can have binding specificities for tryptase and IL-13, tryptase and IL-4, tryptase and IL-5, tryptase and IL-17, or tryptase and IL-33. In particular, the bispecific antibody can have binding specificities for tryptase and IL-13 or tryptase and IL-33. The bispecific antibody can be prepared as a full-length antibody or an antibody fragment.

[0237] For example, in some examples, the bispecific antibody comprises a first binding domain that binds to tryptase and a second binding domain that binds to IL-13. In some embodiments, the first binding domain that binds to tryptase comprises, for example, (a) HVR-H1 comprising the amino acid sequence of X1X2GMX3 (SEQ ID NO: 1) where X1 is Asp or Ser, X2 is Tyr or Phe, and X3 is Val or His; (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of RX1X2X3DWYFDV (SEQ ID NO: 3) where X1 is Asn or Asp, X2 is Tyr or Asn, and X3 is Asp or Tyr; (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5); and (f) at least one, two, three, four, five, or six hypervariable regions (HVRs) selected from HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6), or one or more combinations of the above HVRs, and one or more variants thereof having at least about 80% sequence identity (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 1-6.In some examples, the second binding domain that binds to IL-13 is, for example, (a) HVR-H1 comprising the amino acid sequence of AYSVN (SEQ ID NO: 84), (b) HVR-H2 comprising the amino acid sequence of MIWGDGKIVYNSALKS (SEQ ID NO: 85), (c) HVR-H3 comprising the amino acid sequence of DGYYPYAMDN (SEQ ID NO: 86), (d) HVR-L1 comprising the amino acid sequence of RASKSVDSYGNSFMH (SEQ ID NO: 87), (e) HVR-L2 comprising the amino acid sequence of LASNLES (SEQ ID NO: 88), and (f) HVR-L3 comprising the amino acid sequence of QQNNEDPRT (SEQ ID NO: 89), at least 1, 2, 3, 4, 5, or 6 HVRs selected therefrom, or one or more combinations of the above HVRs, and one or more variants thereof having at least about 80% sequence identity (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 84-89. In some embodiments, the second binding domain comprises 1, 2, 3, 4, 5, or 6 HVRs of anti-IL-13 lebrikizumab.

[0238] For example, in some examples, the first binding domain that binds to trypsin contains at least 1, 2, 3, 4, 5, or 6 hypervariable regions (HVRs) selected from: (a) HVR-H1 containing the amino acid sequence of (a) DYGMV (SEQ ID NO: 7) such as hu31a.v11; (b) HVR-H2 containing the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2); (c) HVR-H3 containing the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8); (d) HVR-L1 containing the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4); (e) HVR-L2 containing the amino acid sequence of RTSDLAS (SEQ ID NO: 5); and (f) HVR-L3 containing the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some examples, the second binding domain that binds to IL-13 may contain at least 1, 2, 3, 4, 5, or 6 HVRs selected from, for example: (a) HVR-H1 containing the amino acid sequence of AYSVN (SEQ ID NO: 84); (b) HVR-H2 containing the amino acid sequence of MIWGDGKIVYNSALKS (SEQ ID NO: 85); (c) HVR-H3 containing the amino acid sequence of DGYYPYAMDN (SEQ ID NO: 86); (d) HVR-L1 containing the amino acid sequence of RASKSVDSYGNSFMH (SEQ ID NO: 87); (e) HVR-L2 containing the amino acid sequence of LASNLES (SEQ ID NO: 88); and (f) HVR-L3 containing the amino acid sequence of QQNNEDPRT (SEQ ID NO: 89). In some embodiments, the second binding domain contains 1, 2, 3, 4, 5, or 6 HVRs of anti-IL-13 lebrikizumab. In some embodiments, the first binding domain contains the amino acid sequence of VH and / or VL of hu31a.v11, and the second binding domain contains the amino acid sequence of VH and / or VL of the anti-IL-13 antibody lebrikizumab.

[0239] Any of the aforementioned bispecific anti-tryptase / anti-IL-13 antibodies may comprise a first binding domain that binds to tryptase and that comprises a VH domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 9, or an amino acid sequence having that sequence, a VL domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 10, or an amino acid sequence having that sequence, or (c) a VH domain as in (a) and a VL domain as in (b). Any of the aforementioned bispecific anti-tryptase / anti-IL-13 antibodies may comprise a second binding domain that binds to IL-13 and that comprises a VH domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 90 or SEQ ID NO: 114, or an amino acid sequence having that sequence, a VL domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 91 or SEQ ID NO: 115, or an amino acid sequence having that sequence, or (c) a VH domain as in (a) and a VL domain as in (b). In some examples, the second binding domain comprises the VH and / or VL domains of rituximab.

[0240] In another embodiment, in some examples, the bispecific antibody comprises a first binding domain that binds to tryptase and a second binding domain that binds to IL-33. The second binding domain that binds to IL-33 can comprise, for example, any of the anti-IL-33 antibodies described in U.S. Patent Publication No. 2016 / 0168242, which is hereby incorporated by reference in its entirety. In some embodiments, the first binding domain that binds to tryptase comprises, for example, (a) HVR-H1 comprising the amino acid sequence of X1X2GMX3 (SEQ ID NO: 1) wherein X1 is Asp or Ser, X2 is Tyr or Phe, and X3 is Val or His; (b) HVR-H2 comprising the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of RX1X2X3DWYFDV (SEQ ID NO: 3) wherein X1 is Asn or Asp, X2 is Tyr or Asn, and X3 is Asp or Tyr; (d) HVR-L1 comprising the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5); and (f) at least 1, 2, 3, 4, 5, or 6 hypervariable regions (HVRs) selected from HVR-L3 comprising the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6), or one or more combinations of the above HVRs, and one or more variants thereof having at least about 80% sequence identity (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 1-6.In some examples, the second binding domain that binds to IL-33 comprises at least 1, 2, 3, 4, 5, or 6 HVRs selected from, for example, (a) HVR-H1 comprising the amino acid sequence of SFSMS (SEQ ID NO: 120), (b) HVR-H2 comprising the amino acid sequence of TISGGKTFTDYVDSVKG (SEQ ID NO: 121), (c) HVR-H3 comprising the amino acid sequence of ANYGNWFFEV (SEQ ID NO: 122), (d) HVR-L1 comprising the amino acid sequence of RASESVAKYGLSLLN (SEQ ID NO: 123), (e) HVR-L2 comprising the amino acid sequence of AASNRGS (SEQ ID NO: 124), and (f) HVR-L3 comprising the amino acid sequence of QQSKEVPFT (SEQ ID NO: 125), or one or more combinations of the above HVRs, and one or more variants thereof having at least about 80% sequence identity (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 120-125. In some embodiments, the second binding domain comprises 1, 2, 3, 4, 5, or 6 HVRs of anti-IL-33 antibody 10C12.38.H6.87Y.58I.

[0241] For example, in some examples, the first binding domain that binds to trypsin contains at least one, two, three, four, five, or six hypervariable regions (HVRs) selected from: (a) HVR-H1 containing the amino acid sequence of DYGMV (SEQ ID NO: 7) such as hu31a.v11; (b) HVR-H2 containing the amino acid sequence of FISSGSSTVYYADTMKG (SEQ ID NO: 2); (c) HVR-H3 containing the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8); (d) HVR-L1 containing the amino acid sequence of SASSSVTYMY (SEQ ID NO: 4); (e) HVR-L2 containing the amino acid sequence of RTSDLAS (SEQ ID NO: 5); and (f) HVR-L3 containing the amino acid sequence of QHYHSYPLT (SEQ ID NO: 6). In some examples, the second binding domain that binds to IL-33 may contain at least one, two, three, four, five, or six HVRs selected from, for example: (a) HVR-H1 containing the amino acid sequence of SFSMS (SEQ ID NO: 120); (b) HVR-H2 containing the amino acid sequence of TISGGKTFTDYVDSVKG (SEQ ID NO: 121); (c) HVR-H3 containing the amino acid sequence of ANYGNWFFEV (SEQ ID NO: 122); (d) HVR-L1 containing the amino acid sequence of RASESVAKYGLSLLN (SEQ ID NO: 123); (e) HVR-L2 containing the amino acid sequence of AASNRGS (SEQ ID NO: 124); and (f) HVR-L3 containing the amino acid sequence of QQSKEVPFT (SEQ ID NO: 125). In some embodiments, the second binding domain contains one, two, three, four, five, or six HVRs of anti-IL-33 antibody 10C12.38.H6.87Y.58I. In some embodiments, the first binding domain contains the amino acid sequence of VH and / or VL of hu31a.v11, and the second binding domain contains the amino acid sequence of VH and / or VL of anti-IL-33 antibody 10C12.38.H6.87Y.58I.

[0242] Any of the aforementioned bispecific anti-tryptase / anti-IL-33 antibodies may comprise a first binding domain that binds to tryptase and includes a VH domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with SEQ ID NO: 9, or an amino acid sequence having that sequence, a VL domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with SEQ ID NO: 10, or an amino acid sequence having that sequence, or (c) a VH domain as in (a) and a VL domain as in (b). Any of the aforementioned bispecific anti-tryptase / anti-IL-33 antibodies may comprise a second binding domain that binds to IL-33 and includes a VH domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with SEQ ID NO: 126, or an amino acid sequence having that sequence, a VL domain comprising an amino acid sequence having at least 80% sequence identity (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with SEQ ID NO: 127, or an amino acid sequence having that sequence, or (c) a VH domain as in (a) and a VL domain as in (b). In some examples, the second binding domain includes the VH and / or VL domains of 10C12.38.H6.87Y.58I.

[0243] Techniques for making multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy-chain-light-chain pairs having different specificities (see Milstein et al. Nature 305:537, 1983, WO93 / 08829, and Traunecker et al. EMBO J. 10:3655, 1991), and "knob-in-hole" engineering (see, e.g., U.S. Patent No. 5,731,168). Multispecific antibodies can also be made by manipulating the electrostatic steering effect to create antibody Fc-heterodimer molecules (WO2009 / 089004A1), cross-linking two or more antibodies or fragments (see, e.g., U.S. Patent No. 4,676,980, and Brennan et al. Science, 229:81, 1985), producing bispecific antibodies using leucine zippers (see, e.g., Kostelny et al. J. Immunol., 148(5):1547-1553, 1992), making bispecific antibody fragments using "diabody" technology (see, e.g., Hollinger et al. Proc. Natl. Acad. Sci. USA 90:6444-6448, 1993), and using single-chain Fv (scFv) dimers (see, e.g., Gruber et al. J. Immunol. 152:5368, 1994), as well as by preparing trispecific antibodies (such as described in Tutt et al. J. Immunol. 147:60, 1991).

[0244] Engineered antibodies having three or more functional antigen-binding sites, including "octopus antibodies" are also included herein (see, e.g., US2006 / 0025576A1).

[0245] The antibodies or fragments herein also include "Dual Acting FAb" or "DAF" that include trypsinase as well as an antigen-binding site that binds to another different antigen (see, e.g., US2008 / 0069820).

[0246] Knobs-into-Holes The use of Knobs-into-Holes as a method for producing multispecific antibodies is described, for example, in U.S. Patent No. 5,731,168, WO2009 / 089004, US2009 / 0182127, US2011 / 0287009, Marvin and Zhu, Acta Pharmacol. Sin. (2005) 26(6):649-658, and Kontermann (2005) Acta Pharmacol. Sin. 26:1-9. A brief non-limiting review is provided below.

[0247] "Knob" refers to at least one amino acid side chain that protrudes from the interface of a first polypeptide so as to stabilize a heteromultimer and thereby preferentially form a heteromultimer over, for example, a homomultimer, and can thus be positioned within a complementary cavity in an adjacent interface (i.e., the interface of a second polypeptide). The knob may be present within the original interface or introduced synthetically (e.g., by changing the nucleic acid encoding the interface). In some embodiments, the nucleic acid encoding the interface of the first polypeptide is changed to encode a knob. To achieve this, the nucleic acid encoding at least one "original" amino acid residue at the interface of the first polypeptide is replaced with a nucleic acid encoding at least one "transferred" amino acid residue having a larger side chain volume than the original amino acid residue. It will be understood that two or more original residues and corresponding transferred residues may be present. The side chain volumes of various amino residues are shown, for example, in Table 1 of US2011 / 0287009 or Table 1 of U.S. Patent No. 7,642,228.

[0248] In some embodiments, the incoming residue for formation of the protrusion is a naturally occurring amino acid residue selected from arginine (R), phenylalanine (F), tyrosine (Y), and tryptophan (W). In some embodiments, the incoming residue is tryptophan or tyrosine. In some embodiments, the prototype residue for formation of the protrusion has a small side-chain volume, such as alanine, asparagine, aspartic acid, glycine, serine, threonine, or valine. See, for example, U.S. Patent No. 7,642,228.

[0249] A "cavity" refers to at least one amino acid side chain that is recessed from the interface of the second polypeptide and thus accommodates a corresponding protrusion on the interface of the adjacent first polypeptide. The cavity may be present within the original interface or may be introduced synthetically (e.g., by changing the nucleic acid encoding the interface). In some embodiments, the nucleic acid encoding the interface of the second polypeptide is modified to encode a cavity. To achieve this, the nucleic acid encoding at least one "original" amino acid residue of the interface of the second polypeptide is replaced with DNA encoding at least one "incoming" amino acid residue having a smaller side-chain volume than the original amino acid residue. It will be understood that two or more original residues and corresponding incoming residues may be present. In some embodiments, the incoming residue for formation of the cavity is a natural amino acid residue selected from alanine (A), serine (S), threonine (T), and valine (V). In some embodiments, the incoming residue is serine, alanine, or threonine. In some embodiments, the prototype residue for formation of the cavity has a large side-chain volume, such as tyrosine, arginine, phenylalanine, or tryptophan.

[0250] The protrusion is "positionable" within the cavity, which means that the protrusion and the spatial position of the cavity at the interface of the first polypeptide and the second polypeptide, as well as the size of the protrusion and the cavity, are such that the protrusion can be located within the cavity without significantly disturbing the normal association of the first polypeptide and the second polypeptide at the interface. Protrusions such as Tyr, Phe, and Trp typically do not extend perpendicular to the axis of the interface and do not have a preferred conformation, so the alignment of the protrusion with the corresponding cavity may, in some cases, depend on modeling of the protrusion / cavity pair based on a three-dimensional structure such as that obtained by X-ray crystallography or nuclear magnetic resonance (NMR). This can be achieved using techniques widely accepted in the art.

[0251] In some embodiments, the knob mutation in the IgG1 constant region is T366W. In some embodiments, the hole mutation in the IgG1 constant region comprises one or more mutations selected from T366S, L368A, and Y407V. In some embodiments, the hole mutation in the IgG1 constant region comprises T366S, L368A, and Y407V.

[0252] In some embodiments, the knob mutation in the IgG4 constant region is T366W. In some embodiments, the hole mutation in the IgG4 constant region comprises one or more mutations selected from T366S, L368A, and Y407V. In some embodiments, the hole mutation in the IgG4 constant region comprises T366S, L368A, and Y407V.

[0253] 7. Antibody Variant In certain embodiments, amino acid sequence variants of the antibodies provided herein are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody, such as inhibitory activity. Amino acid sequence variants of the antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions from, and / or insertions into, and / or substitutions of residues within the amino acid sequence of the antibody. Deletions, insertions, and substitutions can be combined arbitrarily to arrive at the final construct, provided that the final construct possesses the desired characteristics, such as antigen-binding ability.

[0254] a) Substitution, insertion, and deletion variants In certain embodiments, antibody variants having one or more amino acid substitutions are provided. Sites targeted for substitution mutagenesis include the HVRs and FRs. Conservative substitutions are shown in Table 1 under the heading “Preferred Substitutions.” More substantial changes are provided in Table 1 under the heading “Exemplary Substitutions” and are further described below with reference to amino acid side-chain classes. Amino acid substitutions can be introduced into the antibody of interest and the prod...

Claims

1. A medicament for treating tryptase-related lung disorders in a subject in need thereof, comprising an antibody that binds to human tryptase beta 1 or an antigen-binding fragment thereof, wherein the antibody comprises the following six hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of DYGMV (SEQ ID NO: 7), (b) HVR-H2 comprising the amino acid sequence of FISSSGSSVYYADTMKG (SEQ ID NO: 2), (c) HVR-H3 comprising the amino acid sequence of RNYDDWYFDV (SEQ ID NO: 8), (d) HVR-L1 comprising the amino acid sequence of SASSSTYMY (SEQ ID NO: 4), (e) HVR-L2 comprising the amino acid sequence of RTSDLAS (SEQ ID NO: 5), and (f) HVR-L3 comprising the amino acid sequence of QHYH SYPLT (SEQ ID NO: 6).

2. The medicament according to claim 1, wherein the tryptase-related lung disorder is asthma, airway hyperreactivity, or chronic obstructive pulmonary disease (COPD).

3. The medicament according to claim 2, wherein the tryptase-related lung disorder is asthma.

4. The medicament according to claim 1, formulated for administration in combination with a further therapeutic agent.

5. The medicament according to claim 4, wherein the further therapeutic agent is an interleukin-13 (IL-13) axis-binding antagonist, an interleukin-5 (IL-5) axis-binding antagonist, an interleukin-33 (IL-33) axis-binding antagonist, an M1 prime antagonist, an IgE antagonist, a TRPA1 antagonist, a CRTH2 antagonist, a bronchodilator or asthma symptom controller agent, an immunomodulator, a corticosteroid, a Th2 pathway inhibitor, a tyrosine kinase inhibitor, or a phosphodiesterase inhibitor.

6. The medicament according to claim 1, formulated for subcutaneous, intravenous, intramuscular, topical, oral, transdermal, intraperitoneal, intraorbital, implant, inhalation, intrathecal, intravesical, or intranasal administration.

7. The medicament according to claim 1, wherein the antibody comprises (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 9, (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 10, or (c) the VH domain described in (a) and the VL domain described in (b).

8. The pharmaceutical according to claim 1, wherein the VH domain comprises the amino acid sequence of SEQ ID NO:

9.

9. The pharmaceutical according to claim 1, wherein the VL domain comprises the amino acid sequence of SEQ ID NO:

10.

10. The pharmaceutical according to claim 1, wherein the antibody comprises (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 9 and (b) a VL domain comprising the amino acid sequence of SEQ ID NO:

10.

11. The pharmaceutical according to claim 1, wherein the antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76 or SEQ ID NO: 78 and (b) a light chain comprising the amino acid sequence of SEQ ID NO:

77.

12. The pharmaceutical according to claim 1, wherein the antibody is capable of inhibiting the enzymatic activity of human tryptase beta 1.

13. wherein the antibody binds to the tryptase with a K of from about 120 pM to about 0.5 nM Ｄ The pharmaceutical according to claim 12, wherein the antibody binds to the tryptase with a K of from about 120 pM to about 0.5 nM

14. The antibody binds to the tryptase with a K of about 400 pM Ｄ The pharmaceutical according to claim 13, wherein the antibody binds to the tryptase at K Ｄ .

15. The pharmaceutical according to claim 12, wherein the antibody inhibits the activity of the tryptase with an IC50 of about 2.5 nM or less as determined by a human tryptase beta enzyme assay using a chromogenic synthetic peptide substrate.

16. (i) the antibody is capable of inhibiting the enzymatic activity of human tryptase beta 1 at pH 6; (ii) the antibody is capable of inhibiting tryptase-mediated stimulation of bronchial smooth muscle cell proliferation and / or collagen-based contraction; (iii) the antibody is capable of inhibiting mast cell histamine release; (iv) the antibody is capable of inhibiting IgE-induced histamine release and / or tryptase-induced histamine release; (v) the antibody is capable of inhibiting tryptase activity in cynomolgus bronchial alveolar lavage (BAL) or nasal absorption samples; (vi) the antibody is capable of dissociating tetrameric human tryptase beta 1; (vii) the antibody is capable of dissociating tetrameric human tryptase beta 1 when in a monovalent format; and / or (viii) the antibody is capable of dissociating tetrameric human tryptase beta 1 in the presence of heparin, the pharmaceutical according to claim 12.

17. The pharmaceutical according to claim 1, wherein the antibody is capable of dissociating both the small interface of tetrameric human tryptase beta 1 and the large interface of tetrameric human tryptase beta 1.

18. The pharmaceutical according to claim 1, wherein the antibody further binds to cynomolgus tryptase, human tryptase alpha, human tryptase beta 2 and / or human tryptase beta 3.

19. The pharmaceutical according to claim 1, wherein the antibody is humanized.

20. The pharmaceutical according to claim 1, wherein the antibody is an IgG antibody.

21. The pharmaceutical according to claim 20, wherein the IgG antibody is an IgG1 antibody or an IgG4 antibody.

22. The pharmaceutical according to claim 21, wherein the IgG4 antibody contains an S228P mutation (according to EU numbering) in the heavy chain constant region.

23. The pharmaceutical according to claim 21, wherein the antibody is a monospecific antibody.

24. The pharmaceutical according to claim 1, wherein the antibody is a multispecific antibody.

25. The pharmaceutical according to claim 24, wherein the multispecific antibody is a bispecific antibody.

26. The antibody according to claim 25 comprises a first binding domain that binds to human tryptase beta 1 and a second binding domain that binds to a second biological molecule, wherein the second biological molecule is interleukin-13 (IL-13), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-17 (IL-17), IgE, or interleukin-33 (IL-33).

27. The pharmaceutical according to claim 1, wherein the antibody is contained in a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient, or diluent.

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