ASGR inhibitors

Abstract

To provide antigen binding proteins that interact with ASGR, ASGR-1 and / or ASGR-2 as well as methods of making and using such antigen binding proteins.SOLUTION: The present invention provides methods of treating and preventing a cardiovascular disease by administering a pharmaceutically effective amount of ASGR, ASGR-1 and / or ASGR-2 antigen binding proteins. Methods of treating and preventing a cardiovascular disease by administering a pharmaceutically effective amount of interfering RNA compositions that reduce expression of ASGR, ASGR-1 and / or ASGR-2 are described.SELECTED DRAWING: Figure 1A

Description

[Technical field]

[0001] This application claims priority to U.S. Provisional Patent Application No. 62 / 319,740, filed April 7, 2016, U.S. Provisional Patent Application No. 62 / 259,553, filed November 24, 2015, and U.S. Provisional Patent Application No. 62 / 234,546, filed September 29, 2015, which are incorporated by reference in their entireties. References to the Electronic Format of the Sequence Listing and Tables This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on September 28, 2016, is named APMOL017WOSEQUENCE.txt and is 14,772,812 bytes in size.

[0002] Field The field of the invention relates to compositions and methods relating to ASGR inhibitors, including, but not limited to, anti-ASGR, anti-ASGR-1 and / or anti-ASGR-2 antigen binding proteins. [Background technology]

[0003] Cardiovascular disease, which involves the heart or blood vessels, remains the leading cause of worldwide mortality.Cardiovascular disease includes coronary artery disease (CAD), which can cause angina and myocardial infarction (MI), stroke, hypertensive heart disease, rheumatic heart disease and other disorders of the cardiovascular system.Medicines for treating cardiovascular disease, especially coronary artery disease, have been introduced over the past few years (e.g., the small molecule class of drugs called statins, and the recently approved antibody Repatha®, which targets PCSK9). Summary of the Invention [Problem to be solved by the invention]

[0004] Summary of the Invention In some aspects, the present invention provides isolated antigen binding proteins that bind human ASGR and inhibit ASGR function. In one embodiment, the present invention includes isolated antigen binding proteins that bind human ASGR and inhibit ASGR binding to a ligand. In another embodiment, the present invention includes isolated antigen binding proteins that bind human ASGR-1 and inhibit ASGR-1 binding to a ligand and / or ASGR-1 interaction with ASGR-2. In another embodiment, the present invention includes isolated antigen binding proteins that bind human ASGR-2 and inhibit ASGR-2 binding to a ligand and / or ASGR-2 interaction with ASGR-1. In yet another embodiment, the present invention includes isolated antigen binding proteins that bind human ASGR-1 and human ASGR-2 and inhibit ASGR-1 and / or ASGR-2 binding to a ligand. In some embodiments, the isolated binding proteins specifically bind human ASGR, ASGR-1 and / or ASGR-2.

[0005] In some aspects, the invention provides isolated antigen binding proteins that bind human ASGR-1 and that comprise one or more VH CDR1, VH CDR2, or VH CDR3 having an amino acid sequence that is identical to or contains one, two, or three amino acid residue substitutions, deletions, or insertions in each CDR compared to any VH of the sequences set forth in Tables 3-7. In some aspects, the invention includes isolated antigen binding proteins that bind human ASGR-1 and that comprise one or more VL CDR1, VL CDR2, or VL CDR3 having an amino acid sequence that is identical to or contains one, two, or three amino acid residue substitutions, deletions, or insertions in each CDR compared to any VL of the sequences set forth in Tables 3-7. In some embodiments, the isolated antigen binding protein comprises one or more VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set out in Tables 3-7, and one or more VL CDR1, VL CDR2 or VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set out in Tables 3-7. In some embodiments, the isolated antigen binding protein comprises one VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set out in Tables 3-7, and one VL CDR1, VL CDR2 or VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set out in Tables 3-7.In some embodiments, the isolated antigen binding protein comprises two VH CDR1s, VH CDR2s or VH CDR3s having amino acid sequences that are identical to or contain one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any of the VHs of the sequences set out in Tables 3-7, and two VL CDR1s, VL CDR2s or VL CDR3s having amino acid sequences that are identical to or contain one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any of the VLs of the sequences set out in Tables 3-7. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 and a VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set forth in Table 3-7, and a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set forth in Table 3-7. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence that is identical to any of the sequences set forth in Table 3-7. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 or a VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set out in Table A. In some embodiments, the isolated antigen binding protein comprises a VL CDR1, a VL CDR2 or a VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set out in Table A.In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 and a VH CDR3 having an amino acid sequence identical to or containing one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set out in Table A, and a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence identical to or containing one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set out in Table A. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence identical to any of the sequences set out in Table A. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 or a VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set out in Table B. In some embodiments, the isolated antigen binding protein comprises a VL CDR1, a VL CDR2 or a VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set out in Table B. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 and a VH CDR3 having an amino acid sequence identical to or containing one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences depicted in Table B, and a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence identical to or containing one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences depicted in Table B. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence identical to any of the sequences depicted in Table B.Further, in some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 or a VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set out in Table C. In some embodiments, the isolated antigen binding protein comprises a VL CDR1, a VL CDR2 or a VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set out in Table C. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 and a VH CDR3 having an amino acid sequence identical to or containing one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set out in Table C, and a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence identical to or containing one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set out in Table C. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence identical to any of the sequences set out in Table C. In further embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 or a VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set forth in Table 6. In some embodiments, the isolated antigen binding protein comprises a VL CDR1, a VL CDR2 or a VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set forth in Table 6.In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 and a VH CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VH of the sequences set forth in Table 6, and a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence that is identical to or contains one, two or three amino acid residue substitutions, deletions or insertions in each CDR compared to any VL of the sequences set forth in Table 6. In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence that is identical to any of the sequences set forth in Table 6.

[0006] In some aspects, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Tables 3-7. In some aspects, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Tables 3-7. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Tables 3-7 and a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Tables 3-7. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having any of the VH domain amino acid sequences set forth in Tables 3-7, and a light chain variable domain having any of the VL domain amino acid sequences set forth in Tables 3-7. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table A. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table A.In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table A, and a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table A. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having any of the VH domain amino acid sequences set forth in Table A, and a light chain variable domain having any of the VL domain amino acid sequences set forth in Table A. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table B. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table B. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table B, and a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table B. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having any of the VH domain amino acid sequences set forth in Table B, and a light chain variable domain having any of the VL domain amino acid sequences set forth in Table B.In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table C. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table C. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table C, and a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table C. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having any of the VH domain amino acid sequences set forth in Table C, and a light chain variable domain having any of the VL domain amino acid sequences set forth in Table C. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table 6. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 and comprises a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table 6.In some embodiments, the invention provides an isolated antigen binding protein that specifically binds human ASGR-1 and comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table 6, and a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table 6. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds human ASGR-1 and comprises a heavy chain variable domain having any of the VH domain amino acid sequences set forth in Table 6, and a light chain variable domain having any of the VL domain amino acid sequences set forth in Table 6.

[0007] In some aspects, the invention provides an isolated antigen binding protein that binds human ASGR-1 and comprises one or more VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence identical to or containing no more than 18 amino acid residue substitutions, insertions or deletions in each CDR compared to any of the VHs of the sequences set forth in Table 19A as depicted in Figure 55. In some embodiments, the invention provides an isolated antigen binding protein that binds human ASGR-1 and comprises one or more VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence identical to or containing conservative substitutions thereof as any of the amino acid sequences set forth in Table 19B or 19C as depicted in Figure 55. In some aspects, the invention includes an isolated antigen binding protein that binds human ASGR-1 and comprises one or more VL CDR1, VL CDR2, or VL CDR3 having an amino acid sequence that is identical to or contains no more than 14 amino acid residue substitutions, insertions, or deletions in each CDR compared to any of the VLs of the sequences set forth in Table 20A, as depicted in Figure 55. In some embodiments, the invention provides an isolated antigen binding protein that binds human ASGR-1 and comprises one or more VL CDR1, VL CDR2, or VL CDR3 having an amino acid sequence that is identical to or contains conservative substitutions thereof, any of the amino acid sequences set forth in Tables 20B or 20C, as depicted in Figure 55.In some embodiments, the isolated antigen binding protein comprises one or more VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence identical to or containing no more than 18 amino acid residue substitutions, insertions or deletions in each CDR compared to any VH of the sequences set forth in Table 19A as depicted in Figure 55, and one or more VL CDR1, VL CDR2 or VL CDR3 having an amino acid sequence identical to or containing no more than 14 amino acid residue substitutions, insertions or deletions in each CDR compared to any VL of the sequences set forth in Table 20A as depicted in Figure 55. In some embodiments, the invention provides an isolated antigen binding protein that binds to human ASGR-1 and comprises one or more VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence identical to or containing a conservative substitution thereof as set forth in Table 19B or 19C, as depicted in FIG. 55, and one or more VL CDR1, VL CDR2 or VL CDR3 having an amino acid sequence identical to or containing a conservative substitution thereof as set forth in Table 20B or 20C, as depicted in FIG. 55. In some embodiments, the isolated antigen binding protein comprises one VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence identical to or containing no more than 18 amino acid residue substitutions, insertions or deletions in each CDR compared to any VH of the sequences set forth in Table 19A as depicted in Figure 55, and one VL CDR1, VL CDR2 or VL CDR3 having an amino acid sequence identical to or containing no more than 14 amino acid residue substitutions, insertions or deletions in each CDR compared to any VL of the sequences set forth in Table 20A as depicted in Figure 55.In some embodiments, the invention provides an isolated antigen binding protein that binds to human ASGR-1 and comprises one VH CDR1, VH CDR2 or VH CDR3 having an amino acid sequence identical to or containing a conservative substitution thereof as set forth in Tables 19B or 19C, as depicted in FIG. 55, and one VL CDR1, VL CDR2 or VL CDR3 having an amino acid sequence identical to or containing a conservative substitution thereof as set forth in Tables 20B or 20C, as depicted in FIG. 55. In some embodiments, the isolated antigen binding protein comprises two VH CDR1s, VH CDR2s or VH CDR3s having amino acid sequences identical to or containing up to 18 amino acid residue substitutions, insertions or deletions in each CDR compared to any VH of the sequences set forth in Table 19A as depicted in Figure 55, and two VL CDR1s, VL CDR2s or VL CDR3s having amino acid sequences identical to or containing up to 14 amino acid residue substitutions, insertions or deletions in each CDR compared to any VL of the sequences set forth in Table 20A as depicted in Figure 55. In some embodiments, the invention provides an isolated antigen binding protein that binds to human ASGR-1 and comprises two VH CDR1s, VH CDR2s or VH CDR3s having amino acid sequences identical to or containing conservative substitutions of any of the amino acid sequences set forth in Tables 19B or 19C, as depicted in FIG. 55, and two VL CDR1s, VL CDR2s or VL CDR3s having amino acid sequences identical to or containing conservative substitutions of any of the amino acid sequences set forth in Tables 20B or 20C, as depicted in FIG. 55.In some embodiments, the isolated antigen binding protein comprises a VH CDR1, a VH CDR2 and a VH CDR3 having an amino acid sequence identical to or containing up to 18 amino acid residue substitutions, insertions or deletions in each CDR compared to any VH of the sequences set forth in Table 19A as depicted in Figure 55, and a VL CDR1, a VL CDR2 and a VL CDR3 having an amino acid sequence identical to or containing up to 14 amino acid residue substitutions, insertions or deletions in each CDR compared to any VL of the sequences set forth in Table 20A as depicted in Figure 55. In some embodiments, the invention provides an isolated antigen binding protein that binds to human ASGR-1 and comprises a VH CDR1, a VH CDR2 or a VH CDR3 having an amino acid sequence identical to or including a conservative substitution thereof as set forth in Tables 19B or 19C, as depicted in FIG. 55, and a VL CDR1, a VL CDR2 or a VL CDR3 having an amino acid sequence identical to or including a conservative substitution thereof as set forth in Tables 20B or 20C, as depicted in FIG. 55.

[0008] In some aspects, the invention provides isolated antigen binding proteins that specifically bind human ASGR-1 and comprise a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table 19A as depicted in Figure 55, or in Tables 21-34 as depicted in Figure 56, or in Tables 49-95 as depicted in Figure 56. In some aspects, the invention provides isolated antigen binding proteins that specifically bind human ASGR-1 and comprise a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table 20A as depicted in Figure 55, or in Tables 35-48 as depicted in Figure 56, or in Tables 96-134 as depicted in Figure 57. In some embodiments, the antigen binding protein comprises a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences set forth in Table 19A as depicted in Figure 55, or in Tables 21-34 as depicted in Figure 56, or in Tables 49-95 as depicted in Figure 57, and a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences set forth in Table 20A as depicted in Figure 55, or in Tables 35-48 as depicted in Figure 56, or in Tables 96-134 as depicted in Figure 57. In some embodiments, the antigen binding protein comprises a heavy chain variable domain having any of the VH domain amino acid sequences set forth in Table 19A as depicted in Figure 55, or in Tables 21-34 as depicted in Figure 56, or in Tables 49-95 as depicted in Figure 57, and a light chain variable domain having any of the VL domain amino acid sequences set forth in Table 20A as depicted in Figure 55, or in Tables 35-48 as depicted in Figure 56, or in Tables 96-134 as depicted in Figure 57.

[0009] In some aspects, the invention provides an antigen binding protein that specifically binds to human ASGR-1 at an epitope bound by any of the antigen binding proteins disclosed herein. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 at an epitope bound by at least one of the antigen binding proteins set forth in Tables 2-7. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 at an epitope bound by at least one of the antigen binding proteins set forth in Table A. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 at an epitope bound by at least one of the antigen binding proteins set forth in Table B. In some embodiments, the invention provides an isolated antigen binding protein that specifically binds to human ASGR-1 at an epitope bound by at least one of the antigen binding proteins set forth in Table C. In some embodiments, the invention provides isolated antigen binding proteins that specifically bind to human ASGR-1 at an epitope bound by at least one of the antigen binding proteins set forth in Table 6.

[0010] In some aspects, the invention provides isolated antigen binding proteins that compete for binding to human ASGR-1 with any of the antigen binding proteins disclosed herein. In some embodiments, the invention provides isolated antigen binding proteins that compete for binding with any of the antigen binding proteins set forth in Tables 2-7. In some embodiments, the invention provides isolated antigen binding proteins that compete for binding with any of the antigen binding proteins set forth in Table A. In some embodiments, the invention provides isolated antigen binding proteins that compete for binding with any of the antigen binding proteins set forth in Table B. In yet some embodiments, the invention provides isolated antigen binding proteins that compete for binding with any of the antigen binding proteins set forth in Table C. In yet another embodiment, the invention provides isolated antigen binding proteins that compete for binding with any of the antigen binding proteins set forth in Table 6.

[0011] In some aspects, the invention provides isolated antigen binding proteins that bind human ASGR-1 within the carbohydrate recognition domain ("CRD") (also known as the carbohydrate binding domain or "CBD") and inhibit human ASGR-1 binding to a ligand. In some embodiments, the antigen binding proteins bind human ASGR-1 within residues 148-291 or 149-291 or 150-291 or 151-291 or 152-291 or 153-291 or 154-291 or 155-291 of SEQ ID NO:5. In some embodiments, the invention includes isolated antigen binding proteins that bind human ASGR-1 CBD within helix alpha-1. In some embodiments, the invention includes isolated antigen binding proteins that bind human ASGR-1 CBD within residues 174-186 of SEQ ID NO:5. In some embodiments, the invention includes isolated antigen binding proteins that bind human ASGR-1 CBD within helix alpha-2. In some embodiments, the invention includes isolated antigen binding proteins that bind to human ASGR-1 CBD within residues 194-206 of SEQ ID NO:5. In some embodiments, the invention includes isolated antigen binding proteins that bind to human ASGR-1 within residues 237-273 or residues 240-267 of SEQ ID NO:5. In some embodiments, the antigen binding protein binds to ASGR-1 having an amino acid sequence at least 90% identical to SEQ ID NO:5. In some embodiments, the antigen binding protein is an antibody.

[0012] In some aspects, the present invention provides an isolated antigen binding protein or antibody that binds to human ASGR-1 and inhibits human ASGR-1 function. In some embodiments, the isolated antigen binding protein or antibody binds to human ASGR-1 and inhibits human ASGR-1 from binding to a ligand. In some embodiments, the antigen binding protein or antibody or the paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, R237, Q240, D242, W244, E253, N265, D266, D267, N209, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, V268, R271, Y273, N2 09, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, P238, E239, P241, D243, Y245, G246, H247, G252, C255, F258, D260, R263, W264, V268, R271, D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y2 45, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268, C269, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W2 36, D243, E253, F258, G262, W264, D266, H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236,R237, P238, D261, G262, R263, V159, E160, R163, T193, S194, E197, V201, I2 05, G206, P207, Y229, E230, T231, E239, F258, T259, D260, W264, W167, S171 , G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P 272, W275, P155, N157, W158, F168, S169, R170, W175, A178, D179, C182, A187 , W211, C269, R271, Y273, R274, C277, T279, R170, W195, E196, K199, Q202, H 203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261 , G262, R263, R274, N157, V159, F168, S169, S171, S194, Q198, F200, V201, T 210, R237, E239, Q240, F258, T259, W264, H161, S194, W195, E196, Q198, K199 , F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, E160, E162, V 192, T193, E197, V201, H204, Y229, E230, T231, G232, E239, Q240, P241, D26 1, G262, W264, H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E197, Q198, H203, P207, D22 8, E230, W236, R237, D260, G262, or W264, T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, H161, E162, V191, V192, E197, Q198, D216, G219, K222, W2 23, D225, R263, W264, R170, S171, G172, A174, H204, I205, G206, P207, V208,N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W17 5, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269, W275, R1 70, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L 249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264 , D266, V268, C269, W275, N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V20 8, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, R237, Q240, D242, W244, E253, N265, D266, D267, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263,N265, D267, R271, Y273, H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G 262, R263, W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, R170, W195, E196, K199, Q20 2, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, H161, S194, W195, E196, Q198, K199 , F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N 235, P238, D261, R263, T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P2 38, E239, G252, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, R170 , S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R 271, P272, Y273, R274, N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274(SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: Q240, D242, W244,<h2 style=";text-align:left;direction:ltr">E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, R237, Q240, D242, W244 E253, N265, D266, D267, N209, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, V268, R271, Y273, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, P238, E239, P241, D243, Y245, G246, H247, G252, C255, F258, D260, R263, W264, V268, R271, D216, Q217, N218, G219 P220、W221、Y229、E230、K234、W236、E239、Q240、P241、D242、D243、W244、Y2 45、G246、L249、G250、G251、G252、D254、Q270、H215、K222、T231、G232、R237、 P238, H247, G248, E253, C255, D266, V268, C269, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273 Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264, D266, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265 D267, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269, W275 R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, or W275(SE, Q ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, R237, Q240, D242, W244, E253, N265, D266, D267, N209, R237 , Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, D216, Q217, N218, G219, P220, W221, Y 229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D25 4, Q270, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q2 70, R271, P272, Y273, R274, R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, or R274 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, R237, Q240, D242, W244, E253, N265, D266, D267, N209, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, V268, R271, Y273, N209,In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, R237, Q240, D242, W244, E253, N265, D266, D267, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, or Y273 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L24 9, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268, C269, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y2 73, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264, D266, H161, E162, W195, E196, Q198 , K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, V159, E160, R 163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260, W264, W167, S17 1, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, P155, N157, W158,F168, S169, R170, W175, A178, D179, C182, A187, W211, C269, R271, Y273, R2 74, C277, T279, R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207 , V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, N157, V 159, F168, S169, S171, S194, Q198, F200, V201, T210, R237, E239, Q240, F258 , T259, W264, H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K 234, N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204 , Y229, E230, T231, G232, E239, Q240, P241, D261, G262, W264, H161, E162, T 193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261 , R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260, G262, or W264, T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E 230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, :H161, E1 62, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263, W264, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D2 67, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269, W275, R1 70, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246,H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, W275, N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, D244, D245, D246, D247, D248, D249, D250, D251, D252, D253, D254, D255, D256, D257, D258, D259, D260, D261, D262, D263, D264, D265, D266, D267, D268, D269, D270, D271, D272, D273, D274, D275, D276, D277, D278, D279, D280, D281, D282, D283, D284, D285, D286, D287, D288, D289, D290, D291, D292, D293, D294, D295, D296, D297, D298, D299, D300, D301, D302, D303, D304, D305, D306, D307, D308, D309, D310, D311, D312, D313, D314, D315, D316, D317, D318, D319, D320, D321, D322 43, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, W195, N209, N235, R237, P238, E23 9, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R 263, W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P2 72, W275, R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N23 5, W236, P238, D260, D261, G262, R263, R274, H161, S194, W195, E196, Q198, K199, F200, Q202,H203, F233, K234, N235, W236, R237, P238, R263, H161, E162, T193, S194, W195, E19 6, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, T193, S194, W195, E1 96, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R 237, P238, E239, G252, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, R170, S171, G172, K173, A174 , D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T25 9, D260, N265, D267, Q270, R271, P272, Y273, R274, N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, or R274 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268, C269, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264, D266, R170, S171, G172, A174, H204, I205,G206、P207、V208、N209、H257、D260、N265、D267、Q270、R271、P272、Y273、R274、W167、F168、S169、K173、W175、D177、Y181、Q202、H203、T210、W211、R237、F、 258, T259, D261, D266, V268, C269, W275, R170, S171, G172, K173, A174, D177, P207, V2 08, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D26 7, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, or R274 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262,<h2 style=";text-align:left;direction:ltr">R263, V159, E160, R163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260, W264, W167, S171, G172, K173, A174, A176 D177、N180、Y181、R183、L184、E185、D186、Q270、P272、W275、P155、N157、W1 58、F168、S169、R170、W175、A178、D179、C182、A187、W211、C269、R271、Y273、 R274, C277, T279, R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, N157 V159, F168, S169, S171, S194, Q198, F200, V201, T210, R237, E239, Q240, F258, T259, W264, H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233 K234、N235、W236、R237、P238、R263、E160、E162、V192、T193、E197、V201、H2 04、Y229、E230、T231、G232、E239、Q240、P241、D261、G262、W264、H161、E162、 T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260 G262, W264, T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, H161, E162 V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263, W264, N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following residues: H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, W 167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, R170, W195, E1 96, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R27 4, H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, H161, E16 2, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, T193, S194, W195, E196 , P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, or R274 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: H161, E162, W195, E196, Q198, K199, F200, Q202,H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, V159, E160, R163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260 or W264 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262 or R263 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR1 at an epitope that includes at least one of the following amino acid residues: W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, P155, N157, W158, F168, S169, R170, W175, A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, or T279 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272 or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274,N157, V159, F168, S169, S171, S194, Q198, F200, V201, T210, R237, E239, Q240, F258, T259 or W264 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263 or R274 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204, Y229, E230, T231, G232, E239, Q240, P241, D261, G262 or W264 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238 or R263 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260, G262, or W264 (SEQ ID NO:5).The antigen-binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, , F233, K234, N235, P238, D261 or R263 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252,: H161, E162, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263 or W264 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, or G252 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268 or C269 (SEQ ID NO:5).In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254 or Q270 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264 or D266 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271 or Y273 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO:5).In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273 or R274 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269, or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273 or R274 (SEQ ID NO:5).In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E25 3, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269 or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or a paratope in the antibody binds to human ASGR-1 at an epitope that includes at least one of the following amino acid residues: R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273 or R274 (SEQ ID NO:5).

[0013] In some aspects, the present invention provides an isolated antigen binding protein or antibody or a paratope in an antibody that specifically binds to human ASGR-1 and inhibits human ASGR-1 function. In some embodiments, the isolated antigen binding protein or antibody or a paratope in an antibody specifically binds to human ASGR-1 and inhibits human ASGR-1 from binding to a ligand. In some embodiments, the antigen binding protein or antibody or a paratope in an antibody specifically binds to human ASGR-1 within residues 148-291 of SEQ ID NO:5. In some embodiments, when the antigen binding protein or antibody or a paratope in an antibody is bound to human ASGR-1, the antigen binding protein or antibody or a paratope in an antibody is located within 8 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y24 5, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, R237, Q240, D242, W244, E253, N265, D266, D267, N 209, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, V268, R271, Y273, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, P238, E239, P241, D243, Y245, G246, H247, G252, C255, F258, D26 0, R263, W264, V268, R271, D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D2 43, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C 255, D266, V268, C269, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267,R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W2 64, D266, H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232 , F233, K234, N235, W236, R237, P238, D261, G262, R263, V159, E160, R163, T 193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259 , D260, W264, W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L 184, E185, D186, Q270, P272, W275, P155, N157, W158, F168, S169, R170, W175 , A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, T279, R170, W 195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235 , W236, P238, D260, D261, G262, R263, R274, N157, V159, F168, S169, S171, S 194, Q198, F200, V201, T210, R237, E239, Q240, F258, T259, W264, H161, S19 4, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204, Y229, E230, T231, G23 2, E239, Q240, P241, D261, G262, W264, H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E19 7, Q198, H203, P207, D228, E230, W236, R237, D260, G262, or W264, T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233,K234, N235, W236, R237, P238, E239, G252, H161, E162, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R2 63, W264, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272 , Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V 268, C269, W275, R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L24 9, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, W275, N157, R170, S171, G1 72, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, R237, Q240, D242, W244, E253, N265, D266, D267, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273,<h2 style=";text-align:left;direction:ltr">D216、Q217、N218、G219、P220、W221、Y229、E230、K234、W236、E239、Q240、P2 41、D242、D243、W244、Y245、G246、L249、G250、G251、G252、D254、Q270、W195、 N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, H161, E162, W195, E196, Q198, K199, F200, Q202, H203 H204、G232、F233、K234、N235、W236、R237、P238、D261、G262、R263、W167、S1 71、G172、K173、A174、A176、D177、N180、Y181、R183、L184、E185、D186、Q270、 P272, W275, R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, H161, S194 W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233 K234, N235, P238, D261, R263, T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239 G252, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, R170, S171, G172, K173, A174 D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, N157, R170, S171G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, or R274 (SEQ ID NO:5).

[0014] In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, R237, Q240, D242, W244, E253, N265, D266, D267, N209, P238, E239, P241, D243, Y245, G2 46, H247, G252, C255, H257, T259, V268, R271, Y273, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, P238, E239, P241, D243, Y2 45, G246, H247, G252, C255, F258, D260, R263, W264, V268, R271, D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D2 43, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268, C269, W195, N209, N2 35, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W2 64, D266, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W1 75, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268,C269, W275, R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, or W275 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, R237, Q240, D242, W244 , E253, N265, D266, D267, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, D216, Q 217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L24 9, G250, G251, G252, D254, Q270, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N2 65, D267, Q270, R271, P272, Y273, R274, R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, or R274 (SEQ ID NO:5).When the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T 259, D260, V268, R271, Y273, R237, Q240, D242, W244, E253, N265, D266, D267, N209, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, V268, R271, Y273, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, or Y273 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, R237, Q240, D242, W244, E253, N265, D266, D267, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267 or Y273 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266, D267, R237, Q240, D242, W244, E253, N265, D266, D267, N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, or Y273 (SEQ ID NO:5).When bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: Q240, D242, W244, E253, N265, D266 or D267 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: R237, Q240, D242, W244, E253, N265, D266, D267, N209, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, V268, R271 or Y273 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: R237, Q240, D242, W244, E253, N265, D266 or D267 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, P238, E239, P241, D243, Y245, G246, H247, G252, C255, F258, D260, R263, W264, V268 or R271 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266,D267 or Y273 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268 or C269 (see SEQ ID NO: 1). In some embodiments, when the antigen binding protein or antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254 or Q270 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264 or D266 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: W195, N209, N235,In some embodiments, the antigen binding protein or antibody or a paratope in the antibody when bound to human ASGR-1 is an antigen binding tag, The protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269 or W275 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273 or R274 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269 or W275 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: R170, S171,G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273 or R274 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268, C269, W195, N209, N235, R2 37, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264, D266, H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D2 61, G262, R263, V159, E160, R163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260, W264, W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, P155, N157, W158, F168, S169, R170, W1 75, A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, T279, R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207,V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, N157, V1 59, F168, S169, S171, S194, Q198, F200, V201, T210, R237, E239, Q240, F258 , T259, W264, H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K 234, N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204 , Y229, E230, T231, G232, E239, Q240, P241, D261, G262, W264, H161, E162, T 193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261 , R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260, G262, or W264, T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E 230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, H161, E16 2, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263, W264, R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D2 67, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269, W275, R1 70, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y2 73, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245,G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, W275, N157, R170 , S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R27 In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is selected from the group consisting of human ASGR-1 (SEQ ID NO:5). NO:5) located within 5 angstroms of at least one of the following residues: D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q27 0, W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R 271, Y273, H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, W167, S171, G172, K173, A174, A176, D177, N180, Y181, R18 3, L184, E185, D186, Q270, P272, W275, R170, W195, E196, K199, Q202, H203, H204, I205, G206, P2 07, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, H161, S194, W195, E196, Q 198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, H161, E162, T193, S194,W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, T 193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G2 32, F233, K234, N235, W236, R237, P238, E239, G252, R170, S171, G172, A17 4, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274, N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, or R274(SEQ In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, V159, E160, R163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260, W264, W1 67, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, P155, N157, W158, F168, S169, R170, W175, A178, D179, C1 82, A187, W211, C269, R271, Y273, R274, C277, T279, R170, W195, E196, K199,Q202、H203、H204、I205、G206、P207、V208、F233、K234、N235、W236、P238、D260、D261、G262、R263、R274、N157、V159、F168、S169、S171、S194、Q198、F200、V201、 , T210, R237, E239, Q240, F258, T259, W264, H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234 , N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204, Y229, E230, T231, G232, E239, Q240 , P241, D261, G262, W264, H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260, G262, or W264, T193, S194, W 195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G 252, H161, E162, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263, W264, N157, R170, S171, G172, Q 202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO:5).In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P 238, D261, G262, R263, W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, R 170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R2 74, H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, H161, E162, T193, S1 94, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, T193, S194, W195, E196, P220, W221, G226, T22 7, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, N157, R170, S171, G172, Q202, H203, H20 4, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, (SEQ ID NO:5).In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: H161, E162, W195, E196, Q198, K199, F200, Q20 2, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, V159, E160, R163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260 or W264 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues in human ASGR-1 (SEQ ID NO:5): H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262 or R263 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein is located within 8 angstroms of at least one of the following residues: W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, P155, N157, W158, F168, S169, R170, W175, A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, or T279 (SEQ ID NO:5).In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272 or W275 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, N157, V159, F168, S169, S171, S194, Q198, F200, V201, T210, R237, E239, Q240, F258, T259, or W264 (see SEQ ID NO: 1). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the paratope in the antigen binding protein or antibody is located within 5 angstroms of at least one of the following residues: R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, or R274 (SEQ ID NO:5).In some embodiments, when the antigen binding protein or antibody or paratope therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein is located within 8 angstroms of at least one of the following residues: H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204, Y229, E230, T231, G232, E239, Q240, P241, D261, G262 or W264 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238 or R263 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein is located within 8 angstroms of at least one of the following residues: H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260, G262 or W264 (SEQ ID NO:5).In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261 or R263 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein is located within 8 angstroms of at least one of the following residues: T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252,:H161, E162, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263 or W264 (SEQ ID NO:5). In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, or G252 (SEQ ID NO:5).In some embodiments, when the antigen binding protein or antibody or paratope in the antibody is bound to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody is located within 8 angstroms of at least one of the following residues: N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270 or W275 (SEQ ID NO:5). In some embodiments, the antigen binding protein or antibody or paratope in the antibody is related to human ASGR-1. When bound, the antigen binding protein or antibody or paratope in the antibody is located within 5 angstroms of at least one of the following residues: N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273 or R274 (SEQ ID NO:5).

[0015] In some aspects, the present invention includes an isolated antigen binding protein or antibody that specifically binds to human ASGR-1 and inhibits human ASGR-1 function. In some embodiments, the isolated antigen binding protein or antibody that specifically binds to human ASGR-1 inhibits human ASGR-1 binding to a ligand. In some embodiments, the antigen binding protein or antibody specifically binds to human ASGR-1 at a location that overlaps with the location where the ligand binds to human ASGR-1. In some embodiments, the location where the ligand binds to ASGR-1 comprises at least one amino acid residue selected from the group consisting of N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267, Y273, P238, E239, P241, D243, Y245, G246, H247, G252, C255, F258, D260, R263, W264, V268, or R271 (SEQ ID NO:5). In some embodiments, the isolated antigen binding protein or antibody specifically binds to human ASGR-1 at a location that overlaps with the location where the ligand binds to ASGR-1. In some embodiments, the site where the ligand binds to human ASGR-1 includes at least one amino acid residue selected from the group consisting of N209, R237, Q240, D242, W244, E253, H257, T259, N265, D266, D267 and Y273 (SEQ ID NO:5).

[0016] In some aspects, the invention provides an isolated antigen binding protein that binds to human ASGR-1 and inhibits human ASGR, ASGR-1 and / or ASGR-2 function, and does not bind to a variant ASGR-1 protein, wherein the variant ASGR-1 protein has the sequence represented by SEQ ID NO: The present invention includes an antigen binding protein comprising a single mutation at a residue selected from the group consisting of R170, S171, G172, R183, L184, W195, E196, K199, H203, H204, P207, V208, N209, H215, D216, P220, D225, D228, R237, P238, E239, P241, D242, D243, Y245, G246, H247, G248, L249, G251, E253, T259, D260, R263, N265, Q270, R271, P272, R274, and E280, as shown in NO: 5. In some embodiments, an isolated antigen binding protein or antibody is contemplated. An antigen binding protein "does not bind" to a variant ASGR-1 protein if the measured antibody binding signal reduction to the variant ASGR-1 protein (compared to the signal determined for binding to wild-type ASGR-1) is statistically significant as measured by any method known to one of skill in the art, such as the method described in Example 7E below. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of W195, E196, K199, H203, H204, P207, P220, G251 and R263 as shown in SEQ ID NO:5. In some embodiments, the single mutation is selected from the group consisting of H203, H204, P220 and G251. In some embodiments, the single mutation is selected from the group consisting of W195, E196 and K199. In some embodiments, the single mutation is selected from the group consisting of W195, E196 and H204. In some embodiments, the single mutation is selected from the group consisting of W195, K199 and R263. In some embodiments, the single mutation is selected from the group consisting of W195 and E196.In some embodiments, the single mutation is selected from the group consisting of W195 and K199. In some embodiments, the single mutation is selected from the group consisting of W195 or P207. In some embodiments, the single mutation is selected from the group consisting of W195 and R263. In some embodiments, the single mutation is selected from the group consisting of H203 and H204. In some embodiments, the single mutation is selected from the group consisting of K199 and R263. In some embodiments, the single mutation is a mutation of residue W195. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue selected from the group consisting of R170, S171, R183, L184, H215, P220, P238, G246, H247, G248, G251 and N265 as shown in SEQ ID NO:5. In some embodiments, the single mutation is selected from the group consisting of R183, L184, H215, P220, G246, G248, G251 and N265. In some embodiments, the single mutation is selected from the group consisting of L184, P220, P238, H247 and G251. In some embodiments, the single mutation is selected from the group consisting of R170, S171 and L184. In some embodiments, the single mutation is a mutation of residue R183. In some embodiments, the single mutation is a mutation of residue L184. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of P241, D242, D243, Y245, G251, E253 and D260 as shown in SEQ ID NO:5. In some embodiments, the single mutation is selected from the group consisting of P241, D243, Y245, G251, E253 and D260. In some embodiments, the single mutation is selected from the group consisting of P241, D243 and E253. In some embodiments, the single mutation is a mutation at residue D260. In some embodiments, the variant ASGR-1 protein comprises a single mutation at a residue at a position selected from the group consisting of or including R170, R237, E239, P241, T259, D260, R263 and N265 as shown in SEQ ID NO:5.In some embodiments, the single mutation is selected from the group consisting of R237, D260 and R263. In some embodiments, the single mutation is selected from the group consisting of R237, T259, D260 and R263. In some embodiments, the single mutation is selected from the group consisting of R170, R237, P241, T259, D260, R263 and N265. In some embodiments, the single mutation is selected from the group consisting of R237, E239, P241, T259, D260, R263 and N265. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of or including R170, S171, G172, E196, H204, P207, V208, N209, H215, D216, D225, D228, P238, P241, D242, D243, H247, G248, L249, G251, D260, R263, N265, Q270, R271, P272, R274 and E280 as shown in SEQ ID NO:5. In some embodiments, the single mutation is selected from the group consisting of R170, S171, G172, E196, H204, P207, V208, N209, H215, D216, D225, D228, P238, P241, D242, D243, H247, G248, L249, G251, D260, R263, N265, Q270, R271, P272, R274 and E280 as shown in SEQ ID NO:5. In some embodiments, the single mutation is selected from the group consisting of R170, S171, G172, E196, H204, P207, H215, D216, D225, D228, D243, G248, L249, G251, D260, Q270, R271, P272, R274 and E280. In some embodiments, the single mutation is selected from the group consisting of G172, V208, R271, P272 and R274. In some embodiments, the single mutation is selected from the group consisting of G172, R271 and R274. In some embodiments, the single mutation is selected from the group consisting of G172, N209 and R271. In some embodiments, the single mutation is selected from the group consisting of R170, G172, V208, R271 and P272.In some embodiments, the single mutation is selected from the group consisting of G172, V208, P238, R271, P272 and R274. In some embodiments, the single mutation is selected from the group consisting of G172, P238, R271, P272 and R274. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of or including G172, P238, R271 and R274 as shown in SEQ ID NO:5. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of or including R170, G172, V208 and R274 as shown in SEQ ID NO:5. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of or including R170, R183, H215 and Q270 as shown in SEQ ID NO:5. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of or including P241, T259, and N265 as shown in SEQ ID NO:5. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of or including P207 and R263 as shown in SEQ ID NO:5. In some embodiments, the variant ASGR-1 protein comprises a single mutation of a residue at a position selected from the group consisting of or including G172, P241, D242, H247, L249, N265, R271, and P272 as shown in SEQ ID NO:5. In some embodiments, the antigen binding protein or antibody does not bind to more than one variant ASGR-1 protein, which variant ASGR-1 proteins individually comprise a single mutation of this group.

[0017] In some aspects, the invention includes vectors comprising the nucleic acid molecules described herein. In some embodiments, the invention includes host cells comprising the nucleic acid molecules described herein.

[0018] In some aspects, the invention includes nucleic acid molecules encoding the antigen binding proteins described herein.

[0019] In some aspects, the present invention includes pharmaceutical compositions comprising at least one antigen binding protein described herein.

[0020] In some aspects, the present invention provides a method for treating or preventing cardiovascular disease, comprising administering a therapeutically effective dose of an ASGR inhibitor as described herein to a patient in need of cardiovascular disease treatment or prevention. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-2. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1 and ASGR-2. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is one or more of the antigen binding proteins as described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) as described herein. In some embodiments, the relative risk reduction of a cardiovascular event is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60% in a patient.

[0021] In some aspects, the present invention provides a method of reducing the risk of developing coronary artery disease or having a myocardial infarction (MI), comprising administering a therapeutically effective dose of an ASGR inhibitor as described herein to a patient in need of reducing the risk of developing coronary artery disease or having a myocardial infarction (MI). In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-2. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1 and ASGR-2. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is one or more of the antigen binding proteins as described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) as described herein. In some embodiments, the relative risk reduction of coronary artery disease or MI is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60% in a patient.

[0022] In another aspect, the present invention provides a method for lowering blood LDL cholesterol levels in a patient, comprising administering a therapeutically effective dose of an ASGR inhibitor as described herein to a patient in need of lowering blood LDL cholesterol levels. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-2. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1 and ASGR-2. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is one or more of the antigen binding proteins as described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) as described herein. In some embodiments, blood LDL cholesterol is reduced by at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, or at least about 90% compared to pre-dosing levels of blood LDL cholesterol in the patient.

[0023] In yet another aspect, the present invention provides a method for reducing non-HDL cholesterol levels in a patient, comprising administering a therapeutically effective dose of an ASGR inhibitor as described herein to a patient in need of reducing non-HDL cholesterol levels. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-2. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1 and ASGR-2. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is one or more of the antigen binding proteins as described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) as described herein. In some embodiments, non-HDL cholesterol is reduced by at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, or at least about 90% compared to pre-dosing levels of non-HDL cholesterol in the patient.

[0024] In some aspects, the present invention provides a method of increasing alkaline phosphatase ("ALP") levels in a patient, comprising administering to a patient in need of increased ALP levels a therapeutically effective dose of an ASGR inhibitor as described herein. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-2. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1 and ASGR-2. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is one or more of the antigen binding proteins as described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) as described herein. In some embodiments, the ALP level is increased by at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, or at least about 90% compared to the pre-dosing ALP level in the patient. In some embodiments, the ALP level is increased by at least about 1.25x, 1.5x, 2x, 2.5x, 3x, 3.5x, 4x, 4.5x, and 5x compared to pre-treatment.

[0025] In some aspects, the present invention provides a method of antagonizing ASGR, ASGR-1 and / or ASGR-2 in a patient, comprising administering a therapeutically effective dose of an ASGR inhibitor as described herein to a patient in need of antagonizing ASGR-1 and / or ASGR-2. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-2. In some embodiments, the ASGR inhibitor is an inhibitor of ASGR-1 and ASGR-2. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is one or more of the antigen binding proteins as described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) as described herein. [Brief description of the drawings]

[0026] [Figure 1A] FIG. 3 is a diagram of ASGR-1 sequence alignment of human (SEQ ID NO:32699), cyno (SEQ ID NO:32700), dog (SEQ ID NO:32701), pig (SEQ ID NO:32702), rat (SEQ ID NO:32703) and mouse ASGR-1 (SEQ ID NO:32704). Boxed regions indicating different regions of ASGR-1 (i.e., cytoplasmic, transmembrane and carbohydrate binding domains (CBDs; also called carbohydrate binding domains, or CDRs) are representative of the approximate amino acid positions of these regions; human Y273 amino acid is boxed. [Figure 1B] FIG. 1 is a diagram of human ASGR-1 sequence alignment (SEQ ID NOS 32705-32710, respectively, in order of appearance). [Diagram 2]FIG. 3 is a diagram of ASGR-2 sequence alignment for human (SEQ ID NO:32713), cynomolgus monkey (SEQ ID NO:32714), dog (SEQ ID NO:32716), pig (SEQ ID NO:32715), rat (SEQ ID NO:32712) and mouse ASGR-2 (SEQ ID NO:32711). Boxed regions indicating different regions of ASGR-2 (i.e., cytoplasmic, transmembrane and carbohydrate binding domains (CBDs; also called carbohydrate recognition domains or CDRs) are representative of the approximate amino acid positions of these regions. [Diagram 3] FIG. 1 provides an alignment of human ASGR-1 (SEQ ID NO:32717) to human ASGR-2v2 (SEQ ID NO:32718). [Figure 4]Diagram of the del12 variant associated with splicing errors and frameshifts in ASGR-1. (A) Overview of the structure of ASGR-1 mRNA. Exons 4 and 5 are highlighted along with the location of the PCR primers (red arrows) used to amplify the cDNA (the del12 variant is within intron 4 between exons 4 and 5 in the unspliced ​​RNA). (B) Agarose gel showing PCR products generated by amplifying cDNA generated from RNA isolated from blood of del12 carriers and non-carriers. Arrows indicate both the size of the expected PCR product (239 bp) along with the size of the truncated band (217 bp) observed only in del12 heterozygous carriers. (C) Sequence differences between the full-length (239 bp) and variant (217 bp) cDNA fragments based on Sanger sequencing are shown. The variant sequence in del12 carriers is deleted 22 bp at the end of exon 4 compared to the wild-type sequence resulting in a frameshift and introduction of a stop codon. (D) Diagrammatic representation of the splicing defect observed in del12 carriers. Sequences around the exon 4-intron 4 boundary (exon 4 sequence in uppercase and intron 4 sequence in lowercase) are shown along with the 5' splice site in non-carriers and the activated cryptic 5' splice site in del12 carriers. (E) Quantification of full-length (239 bp) and variant (217 bp) cDNA fragments from heterozygous del12 carriers and non-carriers by direct digital counting of sequencing reads generated after sequencing of amplified cDNA products from del12 carriers and non-carriers using the Illumina TruSeq method. The percentage of total ASGR1 transcript incorrectly spliced ​​is shown. Note that the incorrectly spliced ​​form was completely undetectable in non-carriers. [Diagram 5](A) Del12 variants were typed in the indicated populations, totaling 41,648 CAD cases and 247,374 controls. For each cohort, the boxes (diamonds for combined estimates) show the estimated odds ratios, and the lines show the 95% confidence intervals. There was no evidence of heterogeneity across the eight study populations (Phet=0.96). (B) Kaplan-Meier curves for survival against first myocardial infarction in del12 carriers and non-carriers in ASGR-1 stratified by sex. The proportion of individuals who did not have a myocardial infarction is shown on the y-axis and plotted against age on the x-axis. Males and females are represented separately, and a distinction is made in each case between del12 carriers and non-carriers. [Figure 6] Comparison of the association between CAD and non-HDL cholesterol levels between previously identified sequence variants and del12 in ASGR-1. Estimated odds ratio (OR) of the minor allele for coronary artery disease (CAD, 41,648 cases and 247,374 controls) as a function of the estimated effect of the minor allele on non-HDL cholesterol levels (N=119,146) based on the Icelandic population. A complete list of included sequence variants is provided in Table 1.7. Error bars represent 95% confidence intervals. The del12 variant in ASGR-1 is shown. The line indicates the best linear regression fit through the origin. [Figure 7] Analysis of serum ALP, ALT, and AST from ASGR-1 knockout mice is provided. Panel A shows data from male mice studied, and panel B shows data from female mice. [Figure 8]

[0023] Figure 1 shows RNAi in vitro data in CHO cells transfected with hASGR-1 using construct S1662. Panel A is a Western blot demonstrating the reduction in expression of human ASGR-1. Panel B is a graphical representation of the relative reduction in expression of human ASGR-1. Panel C demonstrates that CHO cells receiving construct S1662 show a dramatic reduction in internalization of the ligand (β-GalNAc). [Figure 9]

[0023] Figure 1 shows RNAi in vitro data in CHO cells transfected with mASGR-1 using various constructs. Panel A is a Western blot demonstrating the reduction in expression of mouse ASGR-1. Panel B is a graphical representation of the relative reduction in expression of mouse ASGR-1. Panel C demonstrates that CHO cells receiving various constructs show a dramatic reduction in internalization of the ligand (β-GalNac). [Figure 10]

[0023] Figure 1 shows RNAi in vitro data in HepG2 cells using construct S1662. Panel A is a Western blot demonstrating the reduction of expression of human ASGR-1. Panel B is a graphical representation of the relative reduction of expression of human ASGR-1. [Figure 11] Figure 1 shows RNAi in vitro data in hASGR-2 transfected CHO cells using various constructs. Panel A is a Western blot demonstrating the reduction of human ASGR-2 expression. Panel B is a graphical representation of the relative reduction of human ASGR-2 expression by various constructs. [Figure 12] FIG. 1 shows RNAi in vitro data in CHO cells transfected with mASGR-1 and mASGR-2 using various other constructs. Panel A is a Western blot demonstrating the reduction in expression of mouse ASGR-1 (anti-mouse ASGR-1 or anti-flag) or mouse ASGR-2 (anti-his). Panel B is a graphical representation of the relative reduction in expression of mouse ASGR-1 by various constructs. Panel C is a graphical representation of the relative reduction in expression of mouse ASGR-2 by various constructs. [Figure 13]Figure 1 shows RNAi in vitro data in HepG2 cells using various constructs. Panel A is a Western blot demonstrating the reduction of expression of human ASGR-2. Panel B is a graphical representation of the relative reduction of expression of human ASGR-2 by various constructs. [Figure 14] Figure 1 shows RNAi in vivo data in C57BL / 6J mice using various constructs over a 7 day period, with a total of 3 injections, 1 injection on day 0, 1 injection on day 2, and 1 injection on day 4. Panel A is a graphical representation of quantitative PCR data showing the relative reduction in expression of mASGR-1 RNA in the liver. Panel B is a graphical representation of the relative reduction in expression of mASGR-2 RNA in the liver. [Figure 15] Figure 1 shows RNAi in vivo data in C57BL / 6J mice using various constructs over a 7-day period, with a total of three injections, one on day 0, one on day 2, and one on day 4. Panel A is a Western blot demonstrating the reduction of mouse ASGR-1 protein expression. Panel B is a graphical representation of the relative increase in serum ALP activity. [Figure 16] Figure 1 shows RNAi in vivo data in C57BL / 6J mice using various constructs over 7 days with a single injection on day 0. Panel A is a graphical representation of the relative reduction in expression of mASGR-2 in the liver. Panel B is a graphical representation of the relative reduction in expression of mASGR-1 in the liver. [Figure 17] Figure 1 shows RNAi in vivo data in C57BL / 6J mice with various ASGR-2 constructs over a 7 day period, with a single injection on day 0. This figure is a graphical representation of the relative increase in serum ALP activity. [Figure 18] Panel A shows a computer representation of the crystal structure of the ASGR-1 / lactose complex, Panel B shows a computer representation of the observed electron density, and Panel C shows a close-up of the carbohydrate-binding domain. [Figure 19]Panel A shows a computer representation of the crystal structure of the ASGR-1 / galactose complex, Panel B shows a computer representation of the observed electron density, and Panel C shows a close-up of the carbohydrate-binding domain. [Figure 20] FIG. 11 is a computerized representation of the crystal structure of a close-up of the conformational difference of R237 between the ASGR-1 / lactose (white) and ASGR-1 / galactose (black) complexes. [Figure 21] Panel A shows a computer representation of the crystal structure of the ASGR-1 / GalNAc complex, Panel B shows a computer representation of the observed electron density, and Panel C shows a close-up of the carbohydrate-binding domain. [Figure 22] Panel A shows a structural depiction of the ASGR-1 CBD and 5E5 Fab. Panel B shows a close-up of the ASGR-1 CBD and 5E5 Fab with dashed lines representing the disordered carbohydrate-binding loops and highlighting the ASGR-1 CBD and indirect inhibition of ligand (GalNAc) binding. Panel B also incorporates a double-headed arrow representing the 5 Angstrom distance from tip to tip. [Figure 23] Panel A shows a structural depiction of ASGR-1 CB and 22G5 Fab. Panel B shows a close-up of ASGR-1 CBD and 22G5 Fab with dashed lines representing the disordered carbohydrate-binding loops and highlighting the ASGR-1 CBD and indirect inhibition of ligand (GalNAc) binding. Panel B also incorporates a double-headed arrow representing the 5 angstrom distance from tip to tip. [Figure 24] FIG. 1 is a representation of the structure of ASGR-1 CBD and 4A2 Fab. [Diagram 25] Zoomed-in view of the ASGR-1 CBD and 4A2 Fab structures showing the CDRs of 4A2 Fab interacting with ASGR-1 CBD helix alpha-2 and highlighting indirect inhibition of ASGR-1 CBD and ligand (GalNAc) binding. The view incorporates double arrows representing the 5 angstrom distance from tip to tip. [Figure 26] A close-up of the structure of the ASGR-1 CBD and carbohydrate binding loop with and without the 4A2 Fab, including double arrows representing the 5 angstrom distance from tip to tip. [Figure 27] FIG. 1 is a representation of the structure of ASGR-1 CBD and 7E11 Fab. [Figure 28] A close-up of the structure of the ASGR-1 CBD and 7E11 Fab. The figure depicts the disordered carbohydrate-binding loop with dashed lines, highlighting the indirect inhibition of ASGR-1 CBD and ligand (GalNAc) binding. The figure incorporates double-headed arrows representing the 5 angstrom distance from tip to tip. [Figure 29] FIG. 1 is a representation of the structure of ASGR-1 CBD and 4H6 Fab. [Diagram 30] A close-up of the structure of the ASGR-1 CBD and 4H6 Fab. The figure depicts the disordered carbohydrate-binding loop with dashed lines, highlighting the indirect inhibition of ASGR-1 CBD and ligand (GalNAc) binding. The figure incorporates a double-headed arrow representing a 5 angstrom distance from tip to tip. [Diagram 31] FIG. 1 is a representation of the structure of ASGR-1 CBD and 72G9 Fab. [Diagram 32] Panel A is a close-up of the structure of the ASGR-1 CBD and 72G9 Fab, and panel B is a representation of the structure of the ASGR-1 CBD and 72G9 Fab that also overlaps with the structure of the ASGR-1 CBD and ligand, highlighting direct inhibition of ASGR-1 CBD and ligand (GalNAc) binding. [Diagram 33] FIG. 1 is a representation of the structure of the ASGR-1 CBD and 194A4 Fab. [Diagram 34]A close-up of the structure of the ASGR-1 CBD and 194A4 Fab. The figure depicts the disordered carbohydrate-binding loop with dashed lines, highlighting the indirect inhibition of ASGR-1 CBD and ligand (GalNAc) binding. The figure incorporates a double-headed arrow representing a 5 angstrom distance from tip to tip. [Diagram 35] FIG. 1 is a representation of the structure of ASGR-1 CBD and 54E9 Fab. [Diagram 36] Panel A is a close-up of the structure of the ASGR-1 CBD and 54E9 Fab, and panel B is a representation of the structure of the ASGR-1 CBD and 54E9 Fan that also overlaps with the structure of the ASGR-1 CBD and ligand, highlighting direct inhibition of ASGR-1 CBD and ligand (GalNAc) binding. [Figure 37] Panel A is a representation of the structure of the ASGR-1 CBD and 218G4 Fab, and panel B is an expanded view of the structure of the ASGR-1 CBD and 218G4 Fab. [Figure 38] Panels A and B are close-ups of the structure of 218G4 Fab that also overlap with the structures of the ASGR-1 CBD and the ASGR-1 CBD and ligand. These figures highlight the direct inhibition of ASGR-1 CBD and ligand (GalNAc) binding when 218G4 Fab is present. [Figure 39] FIG. 1 is a representation of the structure of ASGR-1 CBD and 176H4 Fab. [Diagram 40] A close-up of the structure of the 176H4 Fab, which also overlaps with the structures of the ASGR-1 CBD and ASGR-1 and ligand. This figure highlights the direct inhibition of ASGR-1 CBD and ligand (GalNAc) binding when 176H4 Fab is present. [Diagram 41] FIG. 1 is a structural depiction of the ASGR-1 CBD and 194C10 Fab, showing and illustrating the disordered carbohydrate-binding loop with dashed lines, highlighting the ASGR-1 CBD and possible indirect inhibition of ligand (GalNAc) binding. [Diagram 42]Zoomed-in view of the structure of ASGR-1 CBD and 194C10 Fab. This view shows the CDRs of 194C10 that interact with ASGR-1 CBD, highlighting that these may be direct inhibitors of ASGR-1 CBD and ligand (GalNAc) binding. [Diagram 43] Panels AC are graphical representations showing antibody binding results from cells expressing human ASGR-1 and human ASGR-2. [Diagram 44] Panel A is a graphical representation of the effect of the ASGR-1 antibody, 4A2, on serum LDL cholesterol levels in obese cynomolgus monkeys. Panel B is a graphical representation of the effect of the ASGR-1 antibody, 4A2, on serum alkaline phosphatase levels in obese cynomolgus monkeys. Data expressed as % change from baseline. [Diagram 45] Panel A is a graphical representation of the effect of ASGR-1 antibody, 4A2, on serum LDL cholesterol levels in normal cynomolgus monkeys. Panel B is a graphical representation of the effect of ASGR-1 antibody, 4A2, on serum alkaline phosphatase levels in normal cynomolgus monkeys. Data are expressed in percent change from baseline. [Figure 46] Figure 7E shows the coefficient of determination heat map showing the coefficient of determination profile of the test ASGR-1 ligand blocking antibody-reference antibody combinations from arginine / glutamic acid scanning mutagenesis. Dark shading represents highly similar data, whereas light shading represents highly dissimilar data. Relative epitope profiling (antibody competition / binding) bin assignments are also shown. [Figure 47]FIG. 7E is a computer representation showing an alternative view of the ASGR-1 CBD protein and the surface positions of amino acid residues identified by arginine / glutamic acid scanning mutagenesis as important for antibody binding. Relative epitope profiling (antibody competition / binding) bin assignments are also shown. The ligand (GalNAc) is shown as a stick representation (black). The ASGR-1 CBD is shown as a surface representation (light grey). The positions of amino acids identified by Arg / Glu mutation scanning are shown (dark grey surface). The relative positions of important amino acids in each bin are shown for reference only. [Figure 48] 1 is a table presenting various protein sequences for human, mouse, rat, pig, dog and cynomolgus ASGR, ASGR-1 and ASGR-2 (Table 1). [Figure 49] 2A and 2B are two tables presenting the variable light and heavy chain CDR1, CDR2 and CDR3 amino acid sequences for certain antigen binding proteins of the invention. Table 2A presents the variable light chain CDR1, CDR2 and CDR3, while Table 2B presents the variable heavy chain CDR1, CDR2 and CDR3. The CDR sequences in Tables 2A and 2B are folded due to space considerations and should be understood to be single amino acid sequences unless otherwise stated. [Figure 50] A table presenting the amino acid sequences of the light and heavy chain variable domains for certain antigen binding proteins of the invention is shown in Table 3. The amino acid sequences of the light and heavy chain variable domains in Table 3 are folded due to space considerations and should be understood to be single amino acid sequences unless otherwise stated. [Figure 51]4 is a table presenting the protein alignment of the light and heavy variable regions for certain antigen binding proteins of the present invention (Table 4). The asterisk "*" indicates a stop codon. Although sequences containing stop codons are represented as different sequences in the sequence listing, these sequences are related. Generally speaking, however, the amino acid sequences of the light and heavy chain variable domains in the protein alignment presented in Table 4 are folded due to space issues and should be understood to be a single amino acid sequence, unless otherwise stated, as in the case of sequences with one or more stop codons. [Figure 52] 5 is a table presenting the consensus protein alignment of the light and heavy variable regions for certain antigen binding proteins of the present invention (Table 5). The asterisk "*" indicates a stop codon. Sequences containing stop codons are represented in the sequence listing as different sequences, but these sequences are related. Generally speaking, however, the amino acid sequences of the light and heavy chain variable domains in the consensus protein alignment presented in Table 5 are folded due to space issues and should be understood to be a single amino acid sequence, as are sequences with one or more stop codons, unless otherwise stated. [Figure 53] 1 is a table presenting a protein alignment of the light and heavy variable regions for certain optimized antigen binding proteins of the present invention (Table 6). The amino acid sequences of the light and heavy chain variable domains in the protein alignment presented in Table 6 are folded due to space issues and should be understood to be a single amino acid sequence unless otherwise stated. [Figure 54] 7 is a table presenting a consensus protein alignment of the light and heavy variable regions for certain optimized antigen binding proteins of the present invention (Table 7). The amino acid sequences of the light and heavy chain variable domains in the consensus protein alignment presented in Table 7 are folded due to space considerations and should be understood to be single amino acid sequences unless otherwise stated. [Figure 55]

[0023] Figure 19 is a group of tables providing consensus sequences of the various heavy and light chain variable regions (Tables 19A and 20A, respectively) and the CDRs of the various heavy and light chain variable regions (Tables 19B and C and Tables 20B and 20C, respectively) for certain antigen binding proteins of the invention. [Figure 56]

[0023] Figure 21 is a set of tables presenting detailed consensus protein alignments of various light and heavy chain variable regions for certain antigen binding proteins of the present invention (Tables 21-48). The shading of amino acid residues in the consensus protein alignments presented in Tables 21-48 indicates the specific residues that one of skill in the art may wish to target for genetic engineering. [Figure 57] A group of tables providing consensus protein alignments of various light and heavy chain variable regions for certain antigen binding proteins of the present invention (Tables 49-134). [Figure 58] FIG. 1 is a graph showing the reliability of protein measurements in cynomolgus monkeys. The Log10 RFU of mean protein levels in the two species are plotted, with those with low reliability (light dots) and high reliability (darker dots) marked. [Figure 59] Figure 1 shows serum protein analysis of cynomolgus monkeys treated with anti-ASGR-1 antibody. Panel A shows the TNFSF8 protein level in individual animals of different treatment groups over time. Panel B shows the normalized TNFSF8 protein level (percentage of time 0) in individual animals of different treatment groups over time. Panel C shows the TNFSF8 protein level in each treatment group (n=3, error bars represent SEM), and Panel D shows the distribution of TNFSF8 protein level in human ASGR1 del12 carriers and non-carriers. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] As shown in Example 1 below, sequence variants in ASGR-1 (which resulted in either faster degrading ASGR1 or loss-of-function ASGR1 mutations) resulted in reduced levels of non-HDL cholesterol in humans. This in turn resulted in reduced risk of coronary artery disease experienced by such humans. Because loss-of-function mutations in ASGR-1 resulted in both reduced non-HDL cholesterol and reduced coronary artery disease, antibodies and inhibitory RNAs that effectively block ASGR can be used to reduce the risk of coronary artery disease.

[0028] The present invention is directed to inhibitors of ASGR, ASGR-1 and / or ASGR-2. The present invention provides antigen binding proteins that specifically bind to human ASGR, ASGR-1 and / or ASGR-2 and inhibit human ASGR, ASGR-1 and / or ASGR-2 binding to a ligand. The present invention also provides antigen binding proteins that specifically bind to ASGR, ASGR-1 and / or ASGR-2 of other species. The present invention is further directed to a method of treating or preventing cardiovascular disease in a human subject, comprising administering an inhibitor of ASGR, ASGR-1 and / or ASGR-2, wherein the ASGR inhibitor is an antigen binding protein and / or an interfering RNA (e.g., siRNA or shRNA).

[0029] The invention further provides compositions, kits and methods relating to antigen binding proteins that specifically bind to human ASGR, human ASGR-1 and / or human ASGR-2. Nucleic acid molecules comprising a polynucleotide sequence encoding all or part of a polypeptide that specifically binds to human ASGR, human ASGR-1 and / or human ASGR-2 are also provided. The invention further provides vectors and plasmids comprising such nucleic acids, and cells or cell lines comprising such nucleic acids and / or vectors and plasmids. Methods provided further include, for example, methods for making, identifying or isolating antigen binding proteins that bind to human ASGR, human ASGR-1 and / or human ASGR-2, methods for determining whether an antigen binding protein binds to human ASGR, human ASGR-1 and / or human ASGR-2, methods for making compositions, such as pharmaceutical compositions, that comprise antigen binding proteins that bind to human ASGR, human ASGR-1 and / or human ASGR-2, and methods for administering antigen binding proteins that bind to human ASGR, human ASGR-1 and / or human ASGR-2 to a human subject.

[0030] It is to be understood that the foregoing summary and the following detailed description are merely exemplary and explanatory with respect to the invention as claimed, and are not restrictive. In this application, the use of the singular includes the plural, unless specifically stated otherwise. In this application, the use of "or" means "and / or" unless specifically stated otherwise. Furthermore, the use of the term "including" and other forms such as "includes" and "included" is not limiting. Also, terms such as "element" or "component" encompass both elements and components that contain one unit and elements and components that contain two or more subunits, unless specifically stated otherwise. Also, the use of the term "portion" can include a portion of a moiety or the entire moiety.

[0031] Unless otherwise specified herein, scientific and technical terms used in connection with the present invention shall have the meanings commonly understood by those skilled in the art. Furthermore, unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular. In general, the nomenclature used in connection with and techniques of cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are those well known and commonly used in the art. The methods and techniques of the present invention are generally carried out according to conventional methods well known in the art and as described in various general and more detailed references cited and described throughout this specification, unless otherwise specified. See, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989), and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates (1992), and Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1990), which are incorporated herein by reference. Enzymatic reactions and purification techniques are carried out according to manufacturer's specifications, as commonly accomplished in the art or as described herein. The terminology used in connection with analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry, and the laboratory procedures and techniques thereof described herein are well known and commonly used in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and patient treatment.

[0032] Polynucleotide and polypeptide sequences are shown using standard one-letter or three-letter abbreviations. Unless otherwise noted, a polypeptide sequence is oriented with its amino terminus at the left and its carboxy terminus at the right, and single-stranded nucleic acid sequences, and the upper strand of a double-stranded nucleic acid sequence, are oriented with their 5' terminus at the left and their 3' terminus at the right. A particular section of a polypeptide can be designated by amino acid residue number, such as amino acids 1 to 50, or by the actual residue at that site, such as asparagine to proline. A particular polypeptide or polynucleotide sequence can also be described by explaining how it differs from a reference sequence.

[0033] The following terms, unless otherwise indicated, shall be understood to have the following meanings:

[0034] The term "inhibitor" as used herein refers to a compound that reduces the magnitude of at least one activity or function of a molecule compared to the magnitude of the activity or function observed in the absence of the inhibitor. In some cases, an inhibitor substantially reduces the magnitude of at least one activity or function of a molecule compared to the magnitude of the activity or function observed in the absence of the inhibitor. In some cases, an inhibitor completely reduces the magnitude of at least one activity or function of a molecule compared to the magnitude of the activity or function observed in the absence of the inhibitor. Certain exemplary inhibitors include, but are not limited to, proteins, peptides, antibodies, peptibodies, aptamers, antisense oligonucleotides, interfering RNA, carbohydrates, or small organic molecules.

[0035] The term "isolated molecule" (where the molecule is, for example, a polypeptide, polynucleotide, antigen-binding protein, or antibody) is a molecule that, based on its origin or derivation, (1) is not associated with naturally associated components that accompany it in its native state, (2) is substantially free of other molecules from the same species, (3) is expressed by cells from a different species, or (4) does not occur in nature. Thus, a molecule that is chemically synthesized or expressed in a cellular system different from the cell from which it naturally originates, will be "isolated" from its naturally associated components. A molecule may be rendered substantially free of naturally associated components by isolation using purification techniques well known in the art. Molecular purity or homogeneity can be assayed by a number of means well known in the art. For example, the purity of a polypeptide sample can be assayed using polyacrylamide gel electrophoresis and staining of the gel to visualize the polypeptide, using techniques well known in the art. For some purposes, higher resolution may be obtained by using HPLC or other means for purification well known in the art.

[0036] The terms "polynucleotide," "oligonucleotide," and "nucleic acid" are used interchangeably throughout and include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), DNA or RNA analogs produced using nucleotide analogs (e.g., peptide nucleic acids and non-naturally occurring nucleotide analogs), and hybrids thereof. Nucleic acid molecules can be single-stranded or double-stranded. In one embodiment, a nucleic acid molecule of the invention comprises a contiguous open reading frame encoding an antibody of the invention or a fragment, derivative, mutein, or variant thereof.

[0037] A "vector" is a nucleic acid that can be used to introduce another nucleic acid linked to it into a cell. One type of vector is a "plasmid," which refers to a linear or circular double-stranded DNA molecule into which additional nucleic acid segments can be ligated. Another type of vector is a viral vector (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses), in which additional DNA segments can be introduced into the viral genome. Certain vectors can replicate autonomously in a host cell into which they are introduced (e.g., bacterial vectors and episomal mammalian vectors that contain a bacterial origin of replication). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of the host cell after introduction into the host cell, and are thereby replicated along with the host genome. An "expression vector" is a type of vector that can direct the expression of a selected polynucleotide.

[0038] A nucleotide sequence is "operably linked" to a regulatory sequence if the regulatory sequence affects the expression (e.g., the level, timing, or location of expression) of the nucleotide sequence. A "regulatory sequence" is a nucleic acid that affects the expression (e.g., the level, timing, or location of expression) of a nucleic acid to which it is operably linked. A regulatory sequence can exert its effect, for example, directly on the nucleic acid being regulated or through the action of one or more other molecules (e.g., a polypeptide and / or a nucleic acid that binds to the regulatory sequence). Examples of regulatory sequences include promoters, enhancers, and other expression control elements (e.g., polyadenylation signals). Further examples of regulatory sequences are described, for example, in Goeddel, 1990, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA, and Baron et al., 1995, Nucleic Acids Res. 23: 3605-06.

[0039] A "host cell" is a cell that can be used to express a nucleic acid, for example, a nucleic acid of the present invention. A host cell can be a prokaryote, for example, E. coli, or a eukaryote, for example, a unicellular eukaryote (for example, a yeast or other fungus), a plant cell (for example, a tobacco or tomato plant cell), an animal cell (for example, a human cell, a monkey cell, a hamster cell, a rat cell, a mouse cell, or an insect cell), or a hybridoma. Typically, a host cell is a cultured cell that can be transformed or transfected with a polypeptide-encoding nucleic acid, which can then be expressed in the host cell. The phrase "recombinant host cell" can be used to denote a host cell that has been transformed or transfected with a nucleic acid to be expressed. A host cell can also be a cell that contains a nucleic acid but does not express the nucleic acid at a desired level unless a regulatory sequence is introduced into the host cell so that it becomes operably linked to the nucleic acid. It is understood that the term host cell refers not only to the particular subject cell, but also to the progeny or potential progeny of such a cell. Because some modifications may occur in successive generations, due, for example, to mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein. ASGR

[0040] Genomic database analysis is one way that allows the discovery of associations between disease states and specific targets and / or pathways. For example, genetic analysis of familial hypercholesterolemia patients led to the discovery of proprotein convertase subtilisin / kexin type 9 (PCSK9), which is involved in the regulation of serum LDL cholesterol levels and the risk of developing coronary artery disease, and ultimately led to the development of the recently approved anti-hPCSK9 antibody Repatha® (see, for example, Jackson et al., U.S. Patent No. 8,030,457). Advances in DNA sequencing technology provide the means to sequence the genomes of many individuals, allowing the discovery of rare variants. deCODE Genetics (Amgen) previously reported a method to analyze the whole genomes of many Icelanders to explore associations between genetic variants and traits of interest (Gudbjartsson et al., Nature Genetics; Vol. 47; No. (5); May 2015; pp. 435-444).

[0041] This methodology is currently being applied in the search for novel genetic variants that affect cardiovascular disease, including cholesterol levels and the risk of developing coronary artery disease and myocardial infarction (MI). A landmark analysis was performed that identified novel sequence variants in the Ashwell-Morell receptor that are associated with cardiovascular disease.

[0042] In the present invention, whole genome sequencing of Icelanders has discovered a rare 12 base pair deletion ("del12") in intron 4 of the ASGR-1 gene, which is also present in other people of European descent. This deletion results in a frameshift that is predicted to generate a truncated ASGR-1 receptor subunit that lacks both the oligomerization and extracellular carbohydrate recognition domains (also known as "CRDs", "carbohydrate binding domains" or "CBDs"), or may generate an unstable and rapidly degraded transcript (and therefore no protein) by nonsense-mediated decay. In the present invention, whole genome sequencing of Icelanders has also discovered the second rarest loss-of-function variant in the ASGR-1 gene, namely a 4 base pair insertion (c.469-472dupAACT or "W158X") in exon 7. This four base pair insertion in exon 7 causes a frameshift and introduces a premature stop codon at amino acid 158 of the 291 amino acid full-length protein (NP_001662.1:p.W158X). This variant is predicted to code for a protein lacking the carbohydrate recognition domain of the receptor or may generate a transcript that is unstable and rapidly degraded by nonsense-mediated decay (thus resulting in no protein). Furthermore, the W158X variant results in all reported refseq transcripts of ASGR-1, regardless of the tissue or cell type of expression. While not wishing to be constrained by any particular hypothesis, the analysis shows that del12 and W158X result in lower non-HDL cholesterol levels, protection from CAD and MI, and achieves life extension. Moreover, the analysis shows that del12 and W158X are also associated with increased levels of circulating ALP and vitamin B12. Supporting this association of del12 and W158X with increased ALP levels are data from mice carrying the Y272C variant in ASGR-1, which show that such mice display an increased plasma ALP phenotype (Sabrautzki et al., Mamm. Genome, 23, 416-430, 2012).The Y272 position in mouse ASGR-1 corresponds to the Y273 position in human ASGR-1 (see FIG. 1A).

[0043] The Ashwell-Morell receptor (AMR), originally named the hepatic asialoglycoprotein receptor, was one of the first cellular receptors to be isolated and identified (Grewal, Methods in Enzymology, Vol. 479, Chapter 13, 2010, pp. 223-241). This receptor is also known as the Ashwell receptor, the hepatic galactose / N-acetylgalactosamine (GalNAc) receptor, or the hepatic lectin receptor. However, this receptor is now more commonly known as the "ASGPR" or simply "ASGR."

[0044] ASGR is a C-type lectin expressed on the surface of hepatocytes and is composed of a 48 kDa major subunit(s) (ASGR-1) and a 40 kDa minor subunit(s) (ASGR-2) (Roggenbuck et al., Autoimmune Highlights, 2012, 3:119-125). Functional variants of ASGR are formed by oligomerization of the ASGR-1 and ASGR-2 subunits (Grewal). The receptor complex can contain homo- and hetero-oligomers of the ASGR-1 and ASGR-2 subunits, (ASGR-1) 2 -(ASGR-2) 1 The trimer is the most common form and has the highest affinity for substrate (Grewal). Another identified form of ASGR is (ASGR-1). 2 , (ASGR-1) 3 , (ASGR-1) 2 -(ASGR-2) 2 , (ASGR-1) 3 -(ASGR-2) 2 Includes (Grewal). Polynucleotide and polypeptide sequences of ASGR-1 and ASGR-2 of several species are known. Table 1 shows the sequences of human, mouse, rat, pig, dog and cynomolgus monkey. Figure 1A, Figure 1B and Figure 2 show the sequence alignment of ASGR-1 and ASGR-2 of various species, and Figure 3 shows the sequence alignment between human ASGR-1 and human ASGR-2.

[0045] ASGR-1 is a single-pass transmembrane protein and the major subunit of ASGR. The galactose (Gal) or N-acetylgalactosamine (GalNAc) residues of the glycoprotein are exposed by removal of sialic acid by sialidases, and are therefore called asialoglycoproteins for the ligands of ASGR. Although ASGR expression is also detected in other tissues, the liver is the predominant site of expression. A circulating form of the receptor generated from an ASGR-1 transcript lacking exon 2 has also been reported (Liu J, Hu B, Yang Y, et al. A new splice variant of the major subunit of human asialoglycoprotein receptor encodes a secreted form in hepatocytes. PloS one 2010;5:e12934). The del12 and W158X variants are predicted to truncate both the membrane-bound and circulating forms of the receptor, and as described above, the W158X variant may generate transcripts that are unstable and rapidly degraded (and therefore no protein) by nonsense-mediated decay.

[0046] The primary reported function of ASGR is to bind and internalize circulating glycoproteins that contain terminal galactose or N-acetylgalactosamine residues (asialoglycoproteins), resulting in the clearance of these proteins from the circulation (Roggenbuck). Reported endogenous ligands include components of the blood coagulation system, such as platelets and von Willebrand factor (Grewal).

[0047] As used herein, the term "ASGR, ASGR-1 and / or ASGR-2 function" or "ASGR, ASGR-1 and / or ASGR-2 activity" includes any biological effect of ASGR, ASGR-1 and / or ASGR-2. In certain embodiments, ASGR function or activity includes the ability of ASGR to interact or bind with a ligand. In some embodiments, ASGR function or activity is represented by the ability of ASGR to interact or bind with sugars, including but not limited to lactose, galactose and / or GalNAc, or glycoproteins that present such sugars, including but not limited to fetuin, orosomucoid and / or alkaline phosphatase. In some embodiments, ASGR function or activity includes any biological activity resulting from the ASGR response. Exemplary activities include, but are not limited to, clearance of asialoglycoprotein from circulation, clearance of IgA from circulation, removal of apoptotic cells, clearance of low density lipoprotein (LDL) and / or disposal of cellular fibronectin (Roggenbuck).

[0048] Considering the location of ASGR on the surface of hepatocytes in the liver and its significance in hepatocyte entry by certain viruses (Roggenbuck), the receptor has become a convenient target for therapeutic agents that require delivery to the liver and internalization into cells.Examples of such uses include targeted delivery of doxorubicin to hepatocellular carcinoma (Wei et al., Int J Nanomedicine, 2015, 10:5123-37), gene delivery to hepatocytes (D'Souza et al., J Control Release, 2015, 203:126-39) and targeted delivery of siRNA to hepatocytes (Rajeev et al., Chembiochem, 2015, 16(6):903-8).

[0049] Although ASGR and its ability to mediate endocytosis and degradation of desialylated glycoproteins have been known for almost 40 years, the endogenous ligands and physiological functions of this receptor have been difficult to establish (Weigel PH, Yik JH. Glycans as endocytosis signals: the cases of the asialoglycoprotein and hyaluronan / chondroitin sulfate receptors. Biochimica et biophysica acta 2002;1572:341-63). It was reported that ASGR-1- / - mice (lacking any ASGR activity) grow up normally and healthily and do not accumulate desialylated glycoproteins in their circulation, but are unable to clear exogenously added asialoglycoproteins. This suggests that under normal physiological conditions, ASGR is not essential for the homeostasis of circulating asialoglycoproteins (Tozawa R, Ishibashi S, Osuga J, et al. Asialoglycoprotein recaptor deficiency in mice lacking the major receptor subunit. Its obligate requirement for the stable expression of oligomeric receptor. The Journal of Biological Chemistry 2001;276:12624-8).

[0050] In contrast to ASGR-1 knockout mice that lack obvious phenotypes, the present invention establishes the clear physiological role of human ASGR-1 in cardiovascular disease, including but not limited to regulating non-HDL levels and modulating CAD and MI risk.The present invention also demonstrates the association of del12 and W158X with increased circulating ALP and vitamin B12 levels.Furthermore, the present invention shows that disruption of one allele of ASGR-1 appears to have an overall beneficial effect, since del12 heterozygous carriers live an average of 1.5 years longer than non-carriers.

[0051] Surprisingly, various embodiments provided herein demonstrate that both del12 variant and W158 variant have effects on non-HDL levels that are opposite to their effects on ALP and vitamin B12 levels; decreasing non-HDL and increasing ALP and vitamin B12.Without wishing to be bound by any particular hypothesis, it is important to note that the previously described common variants associated with ALP and LDL cholesterol also have opposite effects on these serum components; thus, ASGR-1 may affect the levels of these molecules by different mechanisms.Since other measures of liver function are unaffected, it is unlikely that del12 or W158X mediated ALP increase reflects underlying liver disease. Both circulating ALP and the vitamin B12 transporter, haptocorrin, are nonsialylated glycoproteins known to bind ASGR-1 and be cleared from the circulation by this receptor (Tuin A, Huizinga-Van der Vlag A, van Loenen-Weemaes AM, Meijer DK, Poelstra K. On the role and fate of LPS-dephosphorylating activity in the rat liver. American Journal of Physiology Gastrointestinal and Liver Physiology 2006;290:G377-85; Furger E, Fedosov SN, Lildballe DL, et al. Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin. PloS one 2012;7:e37421; Burger RL, Schneider RJ, Mehlman CS, Allen RH. Human plasma R-type vitamin B12-binding proteins. II.The role of transcobalamin I, transcobalamin III, and the normal granulocyte vitamin B12-binding protein in the plasma transport of vitamin B12., The Journal of Biological Chemistry 1975;250:7707-13; Steirer LM, Park EI, Townsend RR, Baenziger JU., The asialoglycoprotein receptor regulates levels of plasma glycoproteins terminating with sialic acid alpha2,6-galactose., The Journal of Biological Chemistry 2009;284:3777-83). Without wishing to be bound by any particular hypothesis, a more likely basis for the increased levels of ALP and vitamin B12 in del12 and W158X carriers is reduced clearance of desialylated forms of these molecules from the circulation due to a reduced number of functional ASGR receptors in del12 and W158X carriers, suggesting a role for ASGR-1 in maintaining circulating ALP and vitamin B12 homeostasis.

[0052] Without wishing to be bound by any particular hypothesis, the decreased levels of non-HDL in del12 and W158X carriers despite reduced ASGR-1 function suggests that ASGR-1 affects non-HDL levels by mechanisms other than direct binding and endocytosis of cholesterol particles. In mice expressing a hypomorphic form of neuraminidase 1 (Neu1), a sialidase that cleaves sialic acid residues, thereby generating a substrate for ASGR-1, the LDL receptor (LDLR) was sialylated, and this form of the receptor was more stable and took up LDL cholesterol more avidly (LDL levels were reduced in these mice) than the asialylated form of the wild-type LDLR (Yang A, Gyulay G, Mitchell M, White E, Trigatti BL Igdoura SA. Hypomorphic sialidase expression decreases serum cholesterol by downregulation of VLDL production in mice, Journal of Lipid Research 2012;53:2573-2585). Both ASGR and LDLR are located in clathrin-coated pits in hepatocytes, and ASGR may be able to interact with the asialylated form of LDLR and block its activity.

[0053] Two novel rare variants in ASGR-1 are identified herein, which play a role in cardiovascular disease, including but not limited to reducing non-HDL levels and protecting against CAD and MI.These variants disrupt ASGR-1 protein function.Therefore, the present invention is further directed to the method of inhibiting ASGR function, the method of inhibiting ASGR-1 function and / or the method of inhibiting ASGR-2 function.The present invention is further directed to the molecule (including but not limited to, for example, antigen binding protein or interference RNA) that inhibits ASGR function, ASGR-1 function and / or ASGR-2 function.

[0054] Antigen-binding proteins In some embodiments, the invention includes antigen binding proteins that bind to ASGR, ASGR-1 and / or ASGR-2 of different species, including but not limited to human, cynomolgus monkey, pig, dog, mouse and rat. In some embodiments, the antigen binding proteins specifically bind to ASGR, ASGR-1 and / or ASGR-2 of different species, including but not limited to human, cynomolgus monkey, pig, dog and mouse and rat. Exemplary amino acid sequences of human, cynomolgus monkey, dog, pig, rat and mouse ASGR-1 and ASGR-2 are provided in Figures 1-3. In some embodiments, the antigen binding proteins further inhibit ASGR, ASGR-1 and / or ASGR-2 from binding to a ligand.

[0055] An "antigen-binding protein" is a protein that includes an antigen-binding fragment that binds to an antigen and, optionally, a scaffold or framework portion that allows the antigen-binding fragment to adopt a conformation that promotes binding of the antigen-binding protein to the antigen. In this application, the antigen is an ASGR, ASGR-1 and / or ASGR-2 protein, or a fragment thereof. In some embodiments, the antigen-binding fragment includes at least one CDR from an antibody that binds to the antigen, and in some embodiments, includes a heavy chain CDR3 from an antibody that binds to the antigen. In some embodiments, the antigen-binding fragment includes all three CDRs from the heavy chain of the antibody that binds to the antigen or from the light chain of the antibody that binds to the antigen. In some further embodiments, the antigen-binding fragment includes all six CDRs (three from the heavy chain and three from the light chain) from an antibody that binds to the antigen. The antigen-binding fragment is, in certain embodiments, an antibody fragment.

[0056] Non-limiting examples of antigen-binding proteins include antibodies, antibody fragments (e.g., antigen-binding fragments of antibodies), antibody derivatives, and antibody analogs. Further specific examples include, but are not limited to, single chain variable fragments (scFv), nanobodies (e.g., VH domains of camelid heavy chain antibodies; VHH fragments, see Cortez-Retamozo et al., Cancer Research, 64:2853-57, 2004), Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, Fd fragments, and complementarity determining region (CDR) fragments. These molecules can be derived from any mammalian source, such as human, mouse, rat, rabbit, or pig, dog, or camelid. Antibody fragments can compete with intact antibodies for binding to target antigens, and fragments can be produced by modification (e.g., enzymatic or chemical cleavage) of intact antibodies, or can be synthesized de novo using recombinant DNA technology or peptide synthesis. Antigen-binding proteins can include, for example, alternative protein scaffolds or artificial scaffolds with grafted CDRs or CDR derivatives. Such scaffolds include, but are not limited to, antibody-derived scaffolds, including mutations introduced to stabilize the three-dimensional structure of the antigen-binding protein, and fully synthetic scaffolds, including, for example, biocompatible polymers. See, for example, Korndorfer et al., 2003, Proteins: Structure, Function, and Bioinformatics, Vol. 53, No. 1: 121-129 (2003); Roque et al., Biotechnol. Prog., 20: 639-654 (2004). In addition, antibody mimetic-based scaffolds utilizing fibronectin components as scaffolds can be used with peptide antibody mimetics ("PAM").

[0057] Antigen-binding proteins can also include proteins that contain one or more antibody fragments incorporated into a single polypeptide chain or multiple polypeptide chains. For example, antigen-binding proteins can include diabodies (see, e.g., EP 404,097; WO 93 / 11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448, 1993), intrabodies, domain antibodies (a single VL or VH domain or two or more VH domains connected by a peptide linker; see Ward et al., Nature 341:544-546, 1989), maxibodies (two scFvs fused to an Fc region; Fredericks et al., Protein Engineering, Design & Selection 17:95-106, 2004 and Powers et al., Journal of Immunological Sciences 1999). Methods 251:123-135, 2001), triabodies, tetrabodies, minibodies (scFv fused to a CH3 domain; see Olafsen et al., Protein Eng Des Sel. 17:315-23, 2004), peptibodies (one or more peptides attached to an Fc region, see WO00 / 24782), linear antibodies (a pair of tandem Fd segments (VH-CH1-VH-CH1) that, together with complementary light chain polypeptides, form a pair of antigen-binding regions; Zapata et al., Protein Eng Des Sel. 251:123-135, 2001), and the like. Eng. 8:1057-1062, 1995), small modular immunopharmaceuticals (see U.S. Patent Application Publication No. 20030133939), and immunoglobulin fusion proteins (e.g., IgG-scFv, IgG-Fab, 2scFv-IgG, 4scFv-IgG, VH-IgG, IgG-VH and Fab-scFv-Fc).

[0058] In certain embodiments, the antigen-binding protein can have the structure of, for example, an immunoglobulin. An "immunoglobulin" is a tetrameric molecule, with each tetramer containing two identical pairs of polypeptide chains, each pair having one "light" (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain contains a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function.

[0059] Within light and heavy chains, the variable (V) and constant regions (C) are connected by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D" region of about 10 or more amino acids. See generally, Fundamental Immunology, Chapter 7 (Paul, W., ed., 2nd ed., Raven Press, NY (1989)), which is incorporated by reference in its entirety for all purposes. The variable regions of each light / heavy chain pair form the antibody binding site, such that an intact immunoglobulin has two binding sites.

[0060] Immunoglobulin chains exhibit the same general structure of relatively conserved framework regions (FR) connected by three hypervariable regions, also called complementarity determining regions or CDRs. From N- to C-terminus, both light and heavy chains contain the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.

[0061] Human light chains are classified as kappa and lambda light chains. The term "light chain" refers to a polypeptide that includes, from amino to carboxyl terminus, a single immunoglobulin light chain variable region (VL) and a single immunoglobulin light chain constant domain (CL). Heavy chains are classified as mu (μ), delta (Δ), gamma (γ), alpha (α) and epsilon (ε) and define the antibody isotype as IgM, IgD, IgG, IgA and IgE, respectively. The term "heavy chain" refers to a polypeptide that includes, from amino to carboxyl terminus, a single immunoglobulin heavy chain variable region (VH), immunoglobulin heavy chain constant domain 1 (CH1), immunoglobulin hinge region, immunoglobulin heavy chain constant domain 2 (CH2), immunoglobulin heavy chain constant domain 3 (CH3) and optionally immunoglobulin heavy chain constant domain 4 (CH4). The IgG class is further divided into subclasses, namely IgG1, IgG2, IgG3 and IgG4. The IgA class is further divided into subclasses, namely IgA1 and IgA2. IgM has subclasses, including but not limited to IgM1 and IgM2. The heavy chains in IgG, IgA and IgD antibodies have three domains (CH1, CH2 and CH3), while the heavy chains in IgM and IgE antibodies have four domains (CH1, CH2, CH3 and CH4). The immunoglobulin heavy chain constant domains can be from any immunoglobulin isotype, including subtypes. The antibody chains are linked together via interpolypeptide disulfide bonds between the CL and CH1 domains (i.e., between the light and heavy chains) and between the hinge regions of the antibody heavy chains.

[0062] The term "antibody" refers to an intact immunoglobulin of any isotype, including, for example, chimeric, humanized, human, and bispecific antibodies. An "antibody" is a type of antigen-binding protein. An intact antibody generally comprises at least two full-length heavy chains and two full-length light chains. An antibody sequence may be derived exclusively from a single species or may be "chimeric", i.e., different portions of the antibody may be derived from two different species, as described further below. Unless otherwise indicated, the term "antibody" also includes antibodies comprising two substantially full-length heavy chains and two substantially full-length light chains, provided that in this case the antibody retains the same or similar binding and / or function as an antibody composed of two full-length light chains and a heavy chain. For example, antibodies with 1, 2, 3, 4, or 5 amino acid residue substitutions, insertions, or deletions at the N-terminus and / or C-terminus of the heavy and / or light chains are included in this definition, provided that in this case the antibody retains the same or similar binding and / or function as an antibody comprising two full-length heavy chains and two full-length light chains. Further, unless expressly excluded, antibodies include, for example, monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, bispecific antibodies, and synthetic antibodies. In some sections of this disclosure, examples of antigen-binding proteins are described herein in terms of hybridoma lineage number as "number / letter / number" (e.g., 25A4). In such cases, the exact name indicates a specific monoclonal antibody derived from a specific hybridoma having a specific light chain variable region and a heavy chain variable region. In some sections of this disclosure, examples of antigen-binding proteins are described herein in terms of "number / letter / number / 'dot' / number" (e.g., 25A4.001) or "number / letter / number / 'dot' / number / 'dot' / number" (e.g., 25A4.001.001). In such cases, the name indicates a variant of a specific antibody having a light chain variable region and a heavy chain variable region related to but distinct from the antibody derived from the hybridoma.Thus, for example, an antigen binding protein designated 25A4 is not the same as an antibody designated 25A4.001 or 25A4.001.001.

[0063] A "polyclonal antibody" refers to a population of antibodies that typically vary widely in composition and binding specificity. A "monoclonal antibody" ("mAb"), as used herein, refers to one or more of a population of antibodies having identical sequences. A monoclonal antibody binds to an antigen at a specific epitope on the antigen.

[0064] In some embodiments, the antigen-binding protein is an antibody "fragment" or "antigen-binding fragment." As used herein, and unless otherwise specified, "antibody fragment" refers to Fab, Fab', F(ab')2 and Fv fragments that contain at least one CDR of an immunoglobulin sufficient to confer specific antigen binding to ASGR, ASGR-1 and / or ASGR-2. Antibody fragments can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies.

[0065] Fab fragments are monovalent fragments having the VL, VH, CL and CH1 domains; F(ab')2 fragments are bivalent fragments having two Fab fragments linked by disulfide bridges at the hinge regions; Fd fragments have the VH and CH1 domains; Fv fragments have the VL and VH domains of a single arm of an antibody; and dAb fragments have the VH domain, the VL domain or antigen-binding fragments of the VH or VL domains (U.S. Pat. Nos. 6,846,634, 6,696,245; U.S. Pat. Appln. Pub. Nos. 05 / 0202512, 04 / 0202995, 04 / 0038291, 04 / 0009507, 03 / 0039958; Ward et al., Nature 341:544-546 (1989)). In certain embodiments, these antibody fragments may be incorporated into single domain antibodies, single chain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs and bis-scFvs (see, e.g., Hollinger and Hudson, 2005, Nature Biotechnology, 23, 9, pp. 1126-1136). Other antigen-binding proteins contemplated are antibody polypeptides such as those disclosed in U.S. Patent No. 6,703,199, including fibronectin polypeptide monobodies, polypeptides disclosed in U.S. Patent Application Publication No. 2005 / 0238646. In some embodiments, the antibody comprises at least one CDR set forth in Tables 2 or 6 herein.

[0066] A "single-chain variable fragment" ("scFv") is a fusion protein in which the VL and VH domains are connected via a linker (e.g., a synthetic sequence of amino acid residues) to form a contiguous protein chain, which linker is long enough to allow the protein chain to fold back on itself and form a monovalent antigen-binding site (see, e.g., Bird et al., Science 242:423-26 (1988) and Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-83 (1988)). For clarity, a "single-chain variable fragment" is not an antibody or antibody fragment as defined herein. Diabodies are bivalent antibodies that contain two polypeptide chains, each of which contains a VH and VL domain connected by a linker that is too short to allow pairing between the two domains on the same chain, so that each domain is paired with a complementary domain on another polypeptide chain (see, for example, Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-48 (1993) and Poljak et al., Structure 2:1121-23 (1994)). If the two polypeptide chains of a diabody are identical, the diabody resulting from their pairing will have two identical antigen-binding sites. Polypeptide chains with different sequences can be used to create diabodies with two different antigen-binding sites. Similarly, tribodies and tetrabodies are antibodies that contain three and four polypeptide chains, respectively, forming three and four antigen-binding sites, respectively, that can be identical or different.

[0067] The term "CDR" refers to the complementarity determining regions (also called "minimal recognition units" or "hypervariable regions") within an antibody variable sequence. CDRs enable an antigen binding protein to specifically bind to a particular antigen of interest. There are three heavy chain variable region CDRs (CDRH1, CDRH2, and CDRH3) and three light chain variable region CDRs (CDRL1, CDRL2, and CDRL3). The CDRs in each of the two chains are typically aligned by framework regions to form a structure that specifically binds to a specific epitope or domain in the target protein. From the N-terminus to the C-terminus, both naturally occurring light and heavy chain variable regions typically follow the following order of these elements: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. A numbering system has been devised to assign numbers to the amino acids that occupy positions in each of these domains. This numbering system is defined in Kabat Sequences of Proteins of Immunological Interest (1987 and 1991, NIH, Bethesda, MD) or Chothia & Lesk, 1987, J. Mol. Biol. 196:901-917; Chothia et al., 1989, Nature 342:878-883. The complementarity determining regions (CDRs) and framework regions (FRs) of a given antibody can be identified using this system. Other numbering systems for amino acids in immunoglobulin chains include IMGT® (the international ImMunoGeneTics information system; Lefranc et al., Dev. Comp. Immunol. 29:185-203; 2005) and AHo (Honegger and Pluckthun, J. Mol. Biol. 309(3):657-670; 2001). One or more CDRs can be incorporated into a molecule, either covalently or noncovalently, to make the molecule an antigen-binding protein.

[0068] In some embodiments, the antigen-binding proteins of the present invention may incorporate the CDR(s) as part of a larger polypeptide chain, may covalently link the CDR(s) to another polypeptide chain, or may incorporate the CDR(s) non-covalently. An antigen-binding molecule may comprise at least one of the CDRs described herein incorporated into a biocompatible framework structure. In one example, the biocompatible framework structure comprises a polypeptide or portion thereof sufficient to form a conformationally stable structural support or framework or scaffold capable of displaying one or more sequences of antigen-binding amino acids (e.g., CDRs, variable regions, etc.) in localized surface regions. Such structures may be naturally occurring polypeptides or polypeptide "folds" (structural motifs) or may have one or more modifications, such as additions, deletions, or substitutions of amino acids, compared to naturally occurring polypeptides or folds. Such scaffolds may be derived from polypeptides of any species (or more than one species), such as humans, other mammals, other vertebrates, invertebrates, plants, bacteria, or viruses.

[0069] Typically, biocompatible framework structures are based on protein scaffolds or skeletons other than immunoglobulin domains. For example, structures based on fibronectin, ankyrin, lipocalin, neocarzinostain, cytochrome b, CP1 zinc finger, PST1, coiled coil, LACI-D1, Z domain and tendamistat domain may be used (see, for example, Nygren and Uhlen, 1997, Current Opinion in Structural Biology, 7, 463-469).

[0070] An antigen-binding protein can have one or more binding sites. When there are two or more binding sites, the binding sites can be identical or different from each other. For example, an antibody typically has two identical binding sites, but a "bispecific" or "bifunctional" antibody has two different binding sites. The two binding sites of a bispecific antigen-binding protein or antibody bind to two different epitopes that can be present on the same or different protein targets.

[0071] In some embodiments, the ASGR-1 antigen binding protein is a bispecific antibody. In certain embodiments, the bispecific antibody binds to ASGR, ASGR-1 or ASGR-2 and PCSK9. In some embodiments, the bispecific antibody binds to ASGR-1 CBD and inhibits ASGR-1 function in addition to binding to PCSK9 and inhibiting PCSK9 binding to LDLR. Methods for making bispecific antibodies are known in the art. One such method for making "bispecific" or "bifunctional" antigen binding proteins or antibodies involves fusion of hybridomas or linking of Fab' fragments. See, for example, Songsivilai and Lachmann, 1990, Clin. Exp. Immunol. 79:315-321; Kostelny et al., 1992, J. Immunol. 148:1547-1553. Another method involves genetic engineering of the Fc portion of the heavy chain, such as creating "knobs" and "holes" that facilitate heterodimerization of the heavy chains when co-expressed in a cell. US 7,695,963. Yet another method also involves genetic engineering of the Fc portion of the heavy chain, but uses electrostatic steering to promote heterodimerization while preventing homodimerization of the heavy chains when co-expressed in a cell. WO 09 / 089,004, which is incorporated herein by reference in its entirety.

[0072] The term "human antibody" includes antibodies having antibody regions, such as variable and constant regions or domains, that substantially correspond to human germline immunoglobulin sequences known in the art, including, for example, those described by Kabat et al. (1991) (cited above). The human antibodies of the invention can include, for example, in the CDRs, particularly CDR3, amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or in vivo somatic mutation). Human antibodies can have at least one, two, three, four, five or more positions replaced by amino acid residues not encoded by human germline immunoglobulin sequences. The definition of human antibody, as used herein, also contemplates fully human antibodies, including only non-artificially and / or genetically altered human sequences of antibodies, as may be obtained by using techniques or systems known in the art, such as, for example, phage display technology or transgenic mouse technology, including, but not limited to, Xenomouse.

[0073] A humanized antibody has a sequence that differs from that of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and / or additions, such that the humanized antibody is less likely to induce an immune response and / or induces a less severe immune response when administered to a human subject compared to the non-human species antibody. In one embodiment, certain amino acids in the framework and constant domains of the heavy and / or light chains of a non-human species antibody are mutated to produce a humanized antibody. In another embodiment, the constant domain(s) from a human antibody are fused to the variable domain(s) of a non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of a non-human antibody are altered to reduce the immunogenic potential of the non-human antibody when administered to a human subject, where either the altered amino acid residues are not critical to the immunospecific binding of the antibody to the antigen, or the changes made to the amino acid sequence are conservative changes, such that the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to this antigen. Examples of methods for making humanized antibodies can be found in US Pat. Nos. 6,054,297, 5,886,152 and 5,877,293.

[0074] The term "chimeric antibody" refers to an antibody that contains one or more regions from one antibody and one or more regions from one or more other antibodies. In one embodiment, one or more of the CDRs are from a human anti-ASGR, ASGR-1 or ASGR-2 antibody. In another embodiment, all of the CDRs are from a human anti-ASGR, ASGR-1 or ASGR-2 antibody. In another embodiment, CDRs from two or more human anti-ASGR, ASGR-1 or ASGR-2 antibodies are mixed and matched to form a chimeric antibody. For example, a chimeric antibody can include CDR1 from the light chain of a first human anti-ASGR, ASGR-1 or ASGR-2 antibody, CDR2 and CDR3 from the light chain of a second human anti-ASGR, ASGR-1 or ASGR-2 antibody, and CDRs from the heavy chain of a third anti-ASGR, ASGR-1 or ASGR-2 antibody. Furthermore, the framework regions may be derived from one of the same anti-ASGR, ASGR-1 or ASGR-2 antibodies, or from one or more different antibodies, such as a human antibody, or from a humanized antibody. In one example of a chimeric antibody, a portion of the heavy and / or light chain is identical to, homologous to, or derived from an antibody from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical to, homologous to, or derived from an antibody from another species or belonging to another antibody class or subclass. Fragments of such antibodies that exhibit the desired biological activity are also included.

[0075] A "neutralizing antigen-binding protein" or "inhibitory antigen-binding protein" or "antagonizing antigen-binding protein" (e.g., a "neutralizing antibody" or "inhibitory antibody" or "antagonizing antibody") refers to an antigen-binding protein or antibody, respectively, that binds to a target molecule and reduces and / or prevents the biological effect of the target molecule. This can be done, for example, by directly blocking the site on the target molecule where it interacts with other molecules (e.g., blocking the ligand-binding site of a receptor) or by indirectly blocking the site on the target molecule where it interacts with other molecules (such as by structural or energetic changes in the target molecule). In some embodiments, these terms can also refer to an antigen-binding protein or antibody that prevents a target molecule to which it binds from performing a biological function. In assessing the binding and / or specificity of an antigen-binding protein, such as an antibody or an immunologically functional fragment thereof, an antibody or fragment can substantially inhibit the binding of a target molecule to its binding partner if the excess antibody reduces the content of the binding partner bound to the target molecule by at least about 1-20, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-85%, 85-90%, 90-95%, 95-97%, 97-98%, 98-99%, 99.5%, 99.9% and 100%. In some embodiments, the inhibition is complete. Measurement of the reduction in binding can be done using various assays known to those skilled in the art (e.g., in vitro competitive binding assays) and is performed using a relevant control molecule so that actual inhibition is measured. For example, numerous competitive assays are known in the art, including but not limited to competitive ELISA, the use of the BiaCore® platform, the Kinexa® platform, and others.Further examples include solid-phase direct or indirect radioimmunoassays (RA), solid-phase direct or indirect enzyme immunoassays (EIA), sandwich competition assays (see, e.g., Stahli et al., 1983, Methods in Enzymology 9:242-253); solid-phase direct biotin-avidin EIA (see, e.g., Kirkland et al., 1986, J. Immunol. 137:3614-3619); solid-phase direct label assays, solid-phase direct label sandwich assays (see, e.g., Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid-phase direct label RIA using I-125 label (see, e.g., Morel et al., 1988, Molec. Immunol. 25:7-15); solid-phase direct biotin-avidin EIA (see, e.g., Cheung et al., 1990, Virology 176:546-552); and direct labeling RIA (Moldenhauer et al., 1990, Scand. J. Immunol. 32:7-82). Typically, such assays involve the use of purified antigen bound to a solid surface or cells bearing either of these, an unlabeled test antigen binding protein, and a labeled reference antigen binding protein. In some embodiments, in the case of ASGR, ASGR-1 and / or ASGR-2, such neutralizing antigen binding proteins or antibodies can reduce the ability of ASGR, ASGR-1 and / or ASGR-2 to bind to the ligand. In some embodiments, the neutralizing ability is characterized and / or described by a competitive assay. In some embodiments, the neutralizing ability is measured using IC. 50 or EC 50The antigen binding proteins in at least Table C are strong neutralizers. In some embodiments, the antibodies or antigen binding proteins bind to ASGR, ASGR-1 and / or ASGR-2 and neutralize by preventing ASGR, ASGR-1 and / or ASGR-2 from binding to ligands (or by reducing the ability of ASGR, ASGR-1 and / or ASGR-2 to bind to ligands), including sugars such as lactose, galactose and / or GalNAc, or glycoproteins that display such sugars, such as fetuin, orosomucoid and / or alkaline phosphatase.

[0076] Competitive inhibition can be measured by determining the amount of labeled ligand bound to a solid surface or cell in the presence of a test antigen-binding protein. Typically, the test antigen-binding protein is present in excess. Antigen-binding proteins or antibodies identified by competitive assays (competitor antigen-binding proteins or antibodies) include antigen-binding proteins that bind to the same epitope as the reference antigen-binding protein, and antigen-binding proteins that bind to adjacent epitopes that are close enough (sufficiently close to the epitope bound by the reference antigen-binding protein so that steric hindrance occurs) to the epitope bound by the reference antigen-binding protein. Typically, when present in excess, a competitor antigen-binding protein inhibits (e.g., reduces) the specific binding of the reference antigen-binding protein to the target antigen by at least 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 65-70%, 70-75%, or 75% or more. In some embodiments, binding is inhibited by at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%. In some embodiments, binding is inhibited by at least 80-85%, 85-90%, 90-95%, 95-97% or more than 97%, including up to 100% inhibition.

[0077] In some embodiments, a ligand binding assay is used, in which cells expressing target protein (e.g., ASGR-1) are mixed with antigen binding protein, incubated for a certain period of time, and then washed.These cells are then incubated with labeled ligand (e.g., β-GalNAc) for a certain period of time, washed, and analyzed for ligand binding.In this analysis, reduced ligand binding compared to related control antigen binding protein indicates inhibition of binding by antigen binding protein that blocks or inhibits this binding.

[0078] Another way that the reduction in binding can be measured is the half-maximal inhibitory concentration (IC50). IC50 measures the amount or concentration of an antigen-binding protein required to inhibit a given property (e.g., ligand binding) by half. In certain embodiments, the antigen-binding protein (e.g., a human antibody) has an IC50 value of 90 nM or less, in another embodiment, an IC50 value of 80 nM or less, in another embodiment, an IC50 value of 70 nM or less, in another embodiment, an IC50 value of 60 nM or less, in another embodiment, an IC50 value of 50 nM or less, in another embodiment, an IC50 value of 40 nM or less, in another embodiment, an IC50 value of 30 nM or less, and in another embodiment, an IC50 value of 25 nM or less.

[0079] In certain embodiments, the antigen binding proteins of the invention bind to ASGR-1 monomers. In some embodiments, the antigen binding proteins of the invention bind to ASGR-1 oligomers. In further embodiments, the antigen binding proteins of the invention bind to ASGR-2 monomers. In some embodiments, the antigen binding proteins of the invention bind to ASGR-2 oligomers. In certain embodiments, the antigen binding proteins of the invention bind to both ASGR-1 and ASGR-2 monomers. In certain embodiments, the antigen binding proteins of the invention bind to (ASGR-1) 2 -(ASGR-2) 1 In some embodiments, the antigen binding protein of the present invention binds to ASGR oligomers, including trimers. 2In a further embodiment, the antigen binding protein of the present invention binds to ASGR oligomers, including dimers. 3 In yet a further embodiment, the antigen binding protein of the invention binds to ASGR oligomers, including trimers. 2 -(ASGR-2) 2 In a further embodiment, the antigen binding protein of the invention binds to ASGR oligomers, including tetramers. 3 -(ASGR-2) 2 Binds to ASGR oligomers, including pentamers. In some embodiments, the antigen binding proteins of the invention bind to multimeric complexes that include at least two subunits of ASGR-1 and / or ASGR-2.

[0080] In certain embodiments, an antigen binding protein (e.g., an antibody, an antibody fragment, etc.) binds to ASGR, ASGR-1 and / or ASGR-2 and inhibits ASGR, ASGR-1 and / or ASGR-2 from binding to a ligand, and the antigen binding protein contains specific amino acid residues at specific positions in the molecule (e.g., VH, VL or CDR). Such residues may be involved in the binding properties of the desired molecule (e.g., part of the paratope). A "paratope," as used herein, is a location in an antibody that binds to an antigen. A paratope may include some amino acid residues from the VH and / or VL CDRs and may also include residues from the framework regions. A paratope binds to an epitope of an antigen. A paratope may be determined using methodologies similar to those described for epitope determination. Once the amino acid residues responsible for the binding properties of the desired molecule have been identified, this information can be used to design antigen-binding proteins (e.g., antibodies, antibody fragments, etc.) that can bind to ASGR, ASGR-1 and / or ASGR-2 and inhibit ASGR function (e.g., inhibit ASGR, ASGR-1 and / or ASGR-2 from binding to ligand).

[0081] The binding site (or interface) between a representative antibody and human ASGR-1 can be determined / defined in a number of ways. For example, the binding of a representative antigen binding protein (e.g., an antibody) to human ASGR-1 was analyzed using X-ray crystallography in Example 10, and the binding site or interface was determined using distances. The crystal structure of the antibody / huASGR1 complex provides information about which residues of the representative antibody form an interface with human ASGR-1. As mentioned above, one skilled in the art can use this information to design antigen binding proteins and antigen binding protein variants, including those containing variable domains with 90% identity or more, 95% identity or more, 97% identity or more, 99% identity or more, or antigen binding protein variants containing variable domains with 20 or less, 15 or less, or 10 or less, or 5 or less insertions, deletions, and / or substitutions in the light and / or heavy chain variable domains of the antigen binding proteins disclosed herein. It may be desirable to maintain the amino acids in the interface while altering non-interface residues. Thus, in some embodiments, antigen binding proteins and antigen binding protein variants of the antigen binding proteins disclosed herein can be designed and made with one or more amino acid additions, substitutions and / or deletions in one or more CDRs that maintain binding to human ASGR-1 and inhibit ASGR, ASGR-1 and / or ASGR-2 function (e.g., inhibit ASGR, ASGR-1 and / or ASGR-2 from binding to ligand).

[0082] In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or all amino acid residues selected from the group consisting of Q27, R30, D32, H91, Y92, S93, Y94, 12, G28, 129, L33, Q90, P95 and R96 of SEQ ID NO:25010, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or all amino acid residues selected from the group consisting of Q27, R30, D32, H91, Y92, S93, Y94, 12, G28, 129, L33, Q90, P95 and R96 of SEQ ID NO:25010. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or all amino acid residues selected from the group consisting of S30, N31, W52, Y53, D54, S56, N57, Y59, Y101, S102, S103, G104, W105, Y106, D107, Y32, V33, V50, G55, K58, N74, E99, V100 and Y108 of NO:29016. In some embodiments, the light chain variable region comprises at least 1, 2, 3, 4, 5, 6 or all amino acid residues selected from the group consisting of Q27, R30, D32, H91, Y92, S93 and Y94 of SEQ ID NO:25010, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or all amino acid residues selected from the group consisting of S30, N31, W52, Y53, D54, S56, N57, Y59, Y101, S102, S103, G104, W105, Y106 and D107 of SEQ ID NO:29016.In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or all amino acid residues selected from the group consisting of H31, S33, N34, N36, Y38, W56, Y97, Y98, 129, S32, N35, N37, Y55, T59, Q96, N99, T100 of SEQ ID NO:25164, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or all amino acid residues selected from the group consisting of H31, S33, N34, N36, Y38, W56, Y97, Y98, 129, S32, N35, N37, Y55, T59, Q96, N99, T100 of SEQ ID NO:25164. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or all amino acid residues selected from the group consisting of T28, F29, T30, N31, Y32, D33, W50, H52, S55, N57, S99, S100, G101, W102, Y103, Y27, I34, N35, W47, M51, P53, N54, G56, T58, G59, Y104, D106 of NO:29170. In some embodiments, the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7 or all amino acid residues selected from the group consisting of H31, S33, N34, N36, Y38, W56, Y97, Y98 of SEQ ID NO:25164, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or all amino acid residues selected from the group consisting of T28, F29, T30, N31, Y32, D33, W50, H52, S55, N57, S99, S100, G101, W102, Y103 of SEQ ID NO:29170.In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or all amino acid residues selected from the group consisting of I30, Y32, T91, Y92, S93, T94, I96, I2, Q27, N28, I29, S31, L33, N34, T50, S67, Q89, Q90, P95 of SEQ ID NO:24908, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or all amino acid residues selected from the group consisting of I30, Y32, T91, Y92, S93, T94, I96, I2, Q27, N28, I29, S31, L33, N34, T50, S67, Q89, Q90, P95 of SEQ ID NO:24908. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 or all amino acid residues selected from the group consisting of S30, S31, I50, W52, H53, S56, N57, Y59, S01, M102, G103, T28, F29, F32, G33, H35, W47, I51, D54, K58, D99, L100, G104 of NO:28914. In some embodiments, the light chain variable region comprises at least one, two, three, four, five, six or all amino acid residues selected from the group consisting of I30, Y32, T91, Y92, S93, T94, I96 of SEQ ID NO:24908, and / or the heavy chain variable region comprises at least one, two, three, four, five, six, seven, eight, nine, ten or all amino acid residues selected from the group consisting of S30, S31, I50, W52, H53, S56, N57, Y59, S01, M102, G103 of SEQ ID NO:28914.In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or all amino acid residues selected from the group consisting of Y32, S91, Y92, R93, Thr94, Pro95, F97, Ile2, Q27, N28, NAG100, Ile29, S30, S31, Q90 and L96 of SEQ ID NO:24362, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or all amino acid residues selected from the group consisting of Y32, S91, Y92, R93, Thr94, Pro95, F97, Ile2, Q27, N28, NAG100, Ile29, S30, S31, Q90 and L96 of SEQ ID NO:24362. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 or all amino acid residues selected from the group consisting of A33, Val50, Ile51, S52, R53, S54, G55, G56, Y57, Y59, R99, A101, A103, G104, E106, S30, S31, Y32, Met34, N35, W47, S49, Thr58, R72, N74, L100, Val102 and S105 of NO:28368. In some embodiments, the light chain variable region comprises at least 1, 2, 3, 4, 5, 6 or all amino acid residues selected from the group consisting of Y32, S91, Y92, R93, Thr94, Pro95, and F97 of SEQ ID NO:24362, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or all amino acid residues selected from the group consisting of A33, Val50, Ile51, S52, R53, S54, G55, G56, Y57, Y59, R99, A101, A103, G104 and E106 of SEQ ID NO:28368.In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or all amino acid residues selected from the group consisting of Q27, W32, A91, N92, S93, F94, F96, D1, 12, G28, 129, S30, R31, Y49, G50, Q89, Q90 and P95 of SEQ ID NO:24930, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or all amino acid residues selected from the group consisting of Q27, W32, A91, N92, S93, F94, F96, D1, 12, G28, 129, S30, R31, Y49, G50, Q89, Q90 and P95 of SEQ ID NO:24930. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 or all amino acid residues selected from the group consisting of Y33, H35, W50, H52, S55, G57, T58, N59, D99, G100, T101, S102, D31, Y32, L34, W47, I51, N54, G56, Y60, Q65, S103 and F104 of NO:28936. In some embodiments, the light chain variable region comprises at least 1, 2, 3, 4, 5, 6 or all amino acid residues selected from the group consisting of Q27, W32, A91, N92, S93, F94 and F96 of SEQ ID NO:24930, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or all amino acid residues selected from the group consisting of Y33, H35, W50, H52, S55, G57, T58, N59, D99, G100, T101 and S102 of SEQ ID NO:28936.In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all amino acid residues selected from the group consisting of Y32, Y49, T50, Q55, S91, H92, S93, F94, F96, S28, I29, T30, N33, L46, S53, L54, S56, Q89, Q90 and P95 of SEQ ID NO:28074, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all amino acid residues selected from the group consisting of Y32, Y49, T50, Q55, S91, H92, S93, F94, F96, S28, I29, T30, N33, L46, S53, L54, S56, Q89, Q90 and P95 of SEQ ID NO:28074. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 or all amino acid residues selected from the group consisting of G26, F27, T28, S30, S31, Y32, S33, S52, G53, S54, S56, Y57, Y59, R98, G100, S101, R102, V2, F29, N35, S50, T51, S55, I58, R72, G99, G103, F104 and D105 of NO:32080. In some embodiments, the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8 or all amino acid residues selected from the group consisting of Y32, Y49, T50, Q55, S91, H92, S93, F94 and F96 of SEQ ID NO:28074, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or all amino acid residues selected from the group consisting of G26, F27, T28, S30, S31, Y32, S33, S52, G53, S54, S56, Y57, Y59, R98, G100, S101 and R102 of SEQ ID NO:32080.In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or all amino acid residues selected from the group consisting of V29, S30, 132, Y33, L47, Y50, R55, A56, T57, Y94, G28, N31, L48, 149, G51, N54, G58, 159, S68, G69, D93 and S95 of SEQ ID NO:26814, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or all amino acid residues selected from the group consisting of V29, S30, 132, Y33, L47, Y50, R55, A56, T57, Y94, G28, N31, L48, 149, G51, N54, G58, 159, S68, G69, D93 and S95 of SEQ ID NO:26814. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or all amino acid residues selected from the group consisting of V31, Y32, Y33, W50, N52, S55, G57, R98, G99, Y100, D101, I102, T204, V2, Y27, T30, L34, N35, P53, N54, G56, T58, N59, A97, L103 and G105 of NO:30820. In some embodiments, the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or all amino acid residues selected from the group consisting of V29, S30, 132, Y33, L47, Y50, R55, A56, T57 and Y94 of SEQ ID NO:26814, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or all amino acid residues selected from the group consisting of V31, Y32, Y33, W50, N52, S55, G57, R98, G99, Y100, D101, 1102 and T204 of SEQ ID NO:30820. In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region is selected from SEQ ID NO:. 27482; and / or the heavy chain variable region comprises at least one, two, three or all of the amino acid residues selected from the group consisting of N31, Y50, V51, Q54 of SEQ ID NO:27482; Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 or all amino acid residues selected from the group consisting of N30, S31, Y32, S52, Y54, N55, K59, R98, D100, F101, W102, S103, G104, Y105, K107, D110, V2, Y27, T28, F29, G33, W50, A53, G56, N57, H99, Y106 or G108 of NO:31488. In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or all amino acid residues selected from the group consisting of N30, S31, Y32, S52, Y54, N55, K59, R98, D100, F101, W102, S103, G104, Y105, K107 and D110 of SEQ ID NO:31488. In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or all amino acid residues selected from the group consisting of Y33, Y50, D51, N53, K54, S57, V34, S52, R55, P56, G58 and G65 of SEQ ID NO:27780, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or all amino acid residues selected from the group consisting of Y33, Y50, D51, N53, K54, S57, V34, S52, R55, P56, G58 and G65 of SEQ ID NO:27780. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or all amino acid residues selected from the group consisting of Q1, V2, F27, S30, S31, Y32, Y53, D54, W99, Y100, Y101, Y102, G26, T28, F29, G33, W52, G55, R72, N74, N98, Y103, Y104, D107 and V108 of NO:31786.In some embodiments, the light chain variable region comprises at least one, two, three, four, five or all of the amino acid residues selected from the group consisting of Y33, Y50, D51, N53, K54 and S57 of SEQ ID NO:27780, and / or the heavy chain variable region comprises at least one, two, three, four, five, six, seven, eight, nine, ten, eleven or all of the amino acid residues selected from the group consisting of Q1, V2, F27, S30, S31, Y32, Y53, D54, W99, Y100, Y101 and Y102 of SEQ ID NO:31786. In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all amino acid residues selected from the group consisting of H31, G32, D33, G34, K35, Y37, I97, Q98, I99, I2, Q27, S28, L29, L30, T36, E55, Q95, S96, P100 and W101 of SEQ ID NO:26536, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all amino acid residues selected from the group consisting of H31, G32, D33, G34, K35, Y37, I97, Q98, I99, I2, Q27, S28, L29, L30, T36, E55, Q95, S96, P100 and W101 of SEQ ID NO:26536. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or all amino acid residues selected from the group consisting of S31, W52, Y53, D54, Y57, Y59, D102, F103, W104, T28, S30, Y32, G33, W47, I50, I51, S56, K58, Y60, K65, D99, H101, S105 and G106 of NO:30542. In some embodiments, the light chain variable region comprises at least one, two, three, four, five, six, seven, eight or all amino acid residues selected from the group consisting of H31, G32, D33, G34, K35, Y37, I97, Q98 and I99 of SEQ ID NO:26536, and / or the heavy chain variable region comprises at least one, two, three, four, five, six, seven, eight or all amino acid residues selected from the group consisting of S31, W52, Y53, D54, Y57, Y59, D102, F103 and W104 of SEQ ID NO:30542.In some embodiments, the antigen binding protein or antibody comprises a light chain variable region and / or a heavy chain variable region, wherein the light chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all amino acid residues selected from the group consisting of N30, S31, Y33, F50, S54, S68, Y92, E93, W97, S28, V29, G32, L47, G51, A52, S53, R55, A56, G69, Q90, Q91, S94 and S95 of SEQ ID NO:26826, and / or the heavy chain variable region comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all amino acid residues selected from the group consisting of N30, S31, Y33, F50, S54, S68, Y92, E93, W97, S28, V29, G32, L47, G51, A52, S53, R55, A56, G69, Q90, Q91, S94 and S95 of SEQ ID NO:26826. Contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or all amino acid residues selected from the group consisting of R30, Y31, Y33, E50, S54, S56, N58, D98, Y99, G100, S28, Y32, W34, S35, W47, G49, I51, S52, H53, G55, T57, R97, A101, F102 and D103 of NO:30832. In some embodiments, the light chain variable region comprises at least one, two, three, four, five, six, seven, eight or all amino acid residues selected from the group consisting of N30, S31, Y33, F50, S54, S68, Y92, E93 and W97 of SEQ ID NO:26826, and / or the heavy chain variable region comprises at least one, two, three, four, five, six, seven, eight, nine or all amino acid residues selected from the group consisting of R30, Y31, Y33, E50, S54, S56, N58, D98, Y99 and G100 of SEQ ID NO:30832.

[0083] In further embodiments, consensus sequences between the antigen binding proteins of the present invention are envisioned. For example, the variable heavy and variable light chain regions (VH and VL) and CDRs (HCDR1 / 2 / 3 and LCDR1 / 2 / 3) of the present invention comprise consensus sequences derived from a group of related monoclonal antibodies. In some embodiments, antigen binding proteins (e.g., antibodies) may be related by both sequence homology and function. As described herein, a "consensus sequence" refers to an amino acid sequence that has conserved amino acids common among multiple sequences and amino acids that vary within a given amino acid sequence at certain positions. In some embodiments, the amino acid modification at a certain position is a substitution. In some embodiments, the amino acid modification at a certain position is a deletion. In some embodiments, the amino acid modification at a certain position is an addition or insertion. These amino acid modifications will be apparent to one of skill in the art upon analyzing a particular antibody VH, VL and / or CDR sequence.

[0084] For example, antibody sequences were analyzed using the following methodology: the Smith-Waterman algorithm was used to align the amino acid sequences against the translated IMGT germline V, D and J genes. First the V genes were located, then the J genes were located in the region downstream of the located V genes, and finally the D genes were located in the region between the V and J regions. Note that since the D genes are relatively short sequences located in the hypervariable CDR3 region, spurious matching is possible and was taken into account as such.

[0085] Sequences from each group were then subjected to sequence similarity alignment using a program using the standard ClustalW algorithm (see Thompson et al., 1994, Nucleic Acids Res. 22:4673-4680). In some cases, a Biosum cost matrix was used, and a gap creation penalty of 50 was used with a gap extension penalty of 0.1. After the alignment was made and then exported as a PDF image, sequence logos were generated by Geneious (v8.1.7, Biomatters). Consensus sequences were generated in Geneious (v8.1.7, Biomatters) with a 0% threshold and exported as a FASTA file. Amino acids that varied within each group are noted in each consensus sequence with the symbol X. See Table 19A VH consensus 1-14 and Table 20A VL consensus 1-14 in Figure 55 and Table 21-48 in Figure 56 for the consensus sequences resulting from this analysis. In other cases, consensus sequences were generated in Abinitio. For consensus sequences resulting from this analysis, see Table 19A VH consensus-15-60 and Table 20A VL consensus 15-54 in Figure 55 and Table 49-134 in Figure 57.

[0086] Alternatively, different analysis methods may be used that are readily available to those skilled in the art. For example, consensus sequences can be determined using standard phylogenetic analysis of the CDRs corresponding to the VH (i.e., variable heavy, etc.) & VL (i.e., variable light, etc.) of an antibody. For example, the amino acid sequences corresponding to any entire variable domain of a VH or VL can be converted into FASTA format to facilitate processing of comparative alignments and inference of phylogeny. The framework regions of these sequences can then be replaced by artificial linker sequences so that the CDRs alone can be examined without introducing any amino acid position weighting bias due to coincident events (such as unrelated antibodies that happen to share a common germline framework inheritance), while still maintaining the CDRs in close proximity within the same sequence corresponding to the VH or VL. The VH or VL sequences in this format can then be subjected to sequence similarity alignment matching using a program that uses a standard ClustalW-like algorithm (see Thompson et al., 1994, Nucleic Acids Res. 22:4673-4680). A gap creation penalty of 8.0 can be used with a gap extension penalty of 2.0. The program also creates phylogenetic trees (phylogenetic tree diagrams) based on sequence similarity alignments using either UPGMA (unweighted average distance method) or neighbor-joining methods (see Saitou and Nei, 1987, Molecular Biology and Evolution 4:406-425) to construct and illustrate the similarities and differences of sequence groups by branch length comparison and grouping. The original sequence alignments created can be used to empirically test and demonstrate the occurrence of amino acids allowed at each position by the consensus group. A consensus sequence of a group of similar sequences within each CDR can then be prepared.

[0087] In another type of approach, CDR consensus sequences can be determined for each separate CDR, without regard to the sequential association of corresponding VH or VL sequences within the same sequence. In this approach, the consensus sequence can be determined by aligning each H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 in the group, i.e., by aligning the individual H-CDR1 sequences of the antigen binding proteins, the H-CDR1 consensus sequence can be determined, by aligning the individual H-CDR2 sequences of the antigen binding proteins, the H-CDR2 consensus sequence can be determined, by aligning the individual H-CDR3 sequences of the antigen binding proteins, the H-CDR3 consensus sequence can be determined, by aligning the individual L-CDR1 sequences of the antigen binding proteins, the L-CDR1 consensus sequence can be determined, by aligning the individual L-CDR2 sequences of the antigen binding proteins, the L-CDR2 consensus sequence can be determined, and by aligning the individual L-CDR3 sequences of the antigen binding proteins, the L-CDR3 consensus sequence can be determined. Similarities between the sequences within each individual CDR sequence may be identified. Thereby, a consensus sequence of a group of similar sequences within each CDR may be presented.

[0088] Various embodiments of the variable heavy chain (VH) consensus amino acid sequences of the present invention are set forth in Table 19A of FIG. 55 (CDRs are underlined, with CDR1 appearing first). Various embodiments of the VH CDR consensus amino acid sequences of the present invention are set forth in Tables 19B and 19C of FIG. 55. In some cases, an "X" is present in the amino acid sequences set forth in Tables 19A and 19B, indicating that two or more amino acids (or zero amino acids) may be present at this location (see FIG. 56 and FIG. 57 for details of the consensus protein alignment). In some cases, a "-" is present in Table 19A (which is the result of the consensus alignment) indicating that the amino acid is not present at this location (see FIG. 56 and FIG. 57 for details of the consensus protein alignment). The VH consensus sequences and VH CDR consensus sequences are based on the analysis of eight or more aligned VH / VH CDR antibody sequences, as described above. In some cases, the VH / VH CDR consensus sequence is based on an analysis of 25 or more, 50 or more, 75 or more, or 100 or more aligned VH antibody sequences. In one case, the VH / VH CDR consensus sequence is based on an analysis of 149 aligned VH antibody sequences.

[0089] Various embodiments of the variable light (VL) consensus amino acid sequences of the present invention are depicted in Table 20A of FIG. 55 (CDRs are underlined, with CDR1 appearing first). Various embodiments of the VL CDR consensus amino acid sequences of the present invention are depicted in Tables 20B and 20C of FIG. 55. As noted above, in some cases, an "X" is present in the amino acid sequences depicted in Tables 20A and 20B, indicating that two or more amino acids (or zero amino acids) may be present at this location (see FIG. 56 and FIG. 57 for details of the consensus protein alignment). In some cases, a "-" is present in Table 20A (which is the result of the consensus alignment) indicating that no amino acid is present at this location (see FIG. 56 and FIG. 57 for details of the consensus protein alignment). The VL consensus sequences and VL CDR consensus sequences are based on the analysis of eight or more aligned VL / VL CDR antibody sequences, as described above. In some cases, the VL / VL CDR consensus sequence is based on an analysis of 25 or more, 50 or more, 75 or more, or 100 or more, 125 or more, or 150 or more aligned VL antibody sequences. In one case, the VL / VL CDR consensus sequence is based on an analysis of 209 aligned VL antibody sequences.

[0090] As described above, the consensus sequence may, in certain embodiments, include substitutions, deletions, or additions / insertions at different positions in the sequence. Specific examples of such substitutions, deletions, or additions / insertions can be found in Tables 19C and 20C of Figure 55, as well as Tables 21-48 of Figure 56 and Tables 49-134 of Figure 57, all of which are incorporated herein. However, in no way should the amino acid substitutions, deletions, or additions / insertions illustrated in Tables 19A-C and 20A-C of Figure 55 or Tables 21-48 of Figure 56 or Tables 49-134 of Figure 57 be construed as limiting the invention to only amino acid substitutions, deletions, or additions at any position of the identified consensus sequence (VH, VL, and / or CDR) with any amino acid contemplated herein.

[0091] In certain embodiments, antigen binding proteins of the invention comprise three VH CDRs and three VL CDRs, where at least one VH CDR is a VH1 CDR selected from Table 19B or Table 19C, as depicted in Figure 55. In certain embodiments, antigen binding proteins of the invention comprise three VH CDRs and three VL CDRs, where at least one VH CDR is a VH2 CDR selected from Table 19B or Table 19C, as depicted in Figure 55. In certain embodiments, antigen binding proteins of the invention comprise three VH CDRs and three VL CDRs, where at least one VH CDR is a VH3 CDR selected from Table 19B or Table 19C, as depicted in Figure 55. In certain embodiments, an antigen binding protein of the invention comprises three VH CDRs and three VL CDRs, where the VH1 CDRs, VH2 CDRs and VH3 CDRs are selected from Table 19B or Table 19C, as depicted in Figure 55.

[0092] In certain embodiments, antigen binding proteins of the invention comprise three VH CDRs and three VL CDRs, where at least one VL CDR is a VL1 CDR selected from Table 20B or Table 20C, as depicted in Figure 55. In certain embodiments, antigen binding proteins of the invention comprise three VH CDRs and three VL CDRs, where at least one VL CDR is a VL2 CDR selected from Table 20B or Table 20C, as depicted in Figure 55. In certain embodiments, antigen binding proteins of the invention comprise three VH CDRs and three VL CDRs, where at least one VL CDR is a VL3 CDR selected from Table 20B or Table 20C, as depicted in Figure 55. In certain embodiments, an antigen binding protein of the invention comprises three VH CDRs and three VL CDRs, where the VL1 CDR, VL2 CDR and VL3 CDR are selected from Table 20B or Table 20C, as depicted in Figure 55.

[0093] In some embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH. In some embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH1 CDR. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH2 CDR. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH3 CDR. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL1 CDR. In a further embodiment, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL2 CDR.In a further embodiment, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL3 CDR.

[0094] In some embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH consensus sequence. In some embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL consensus sequence. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH1 CDR consensus sequence. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH2 CDR consensus sequence. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VH3 CDR consensus sequence. In further embodiments, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL1 CDR consensus sequence. In a further embodiment, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL2 CDR consensus sequence. In a further embodiment, the antigen binding protein comprises no more than 1, 2, 3, 4, 5 or 6 amino acid additions, deletions or substitutions in the VL3 CDR consensus sequence.

[0095] In some embodiments, framework consensus sequences are encompassed by the present invention. Examples of such framework consensus sequences and additions, deletions or substitutions are shown in Tables 21-48 of Figure 56 and Tables 49-134 of Figure 57 herein.

[0096] In further embodiments, the antigen binding proteins of the present invention bind to ASGR of different species, including but not limited to human, cynomolgus monkey, pig, dog, mouse and rat. In some embodiments, the antigen binding proteins of the present invention bind to human. In some embodiments, the antigen binding proteins of the present invention bind to cynomolgus monkey ASGR. In some embodiments, the antigen binding proteins of the present invention bind to pig ASGR. In some embodiments, the antigen binding proteins of the present invention bind to dog ASGR. In some embodiments, the antigen binding proteins of the present invention bind to mouse ASGR. In some embodiments, the antigen binding proteins of the present invention bind to rat ASGR. In some embodiments, the antigen binding proteins specifically bind to ASGR of different species.

[0097] In some embodiments, the antigen binding proteins of the invention bind to ASGR-1 of different species, including but not limited to human, cynomolgus monkey, pig, dog, mouse and rat. In some embodiments, the antigen binding proteins of the invention bind to human ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to cynomolgus monkey ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to pig ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to dog ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to mouse ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to rat ASGR-1. In some embodiments, the antigen binding proteins specifically bind to ASGR-1 of different species.

[0098] In some embodiments, the antigen binding proteins of the invention bind to ASGR-2 of different species, including but not limited to human, cynomolgus monkey, pig, dog, mouse and rat. In some embodiments, the antigen binding proteins of the invention bind to human ASGR-2. In some embodiments, the antigen binding proteins of the invention bind to cynomolgus monkey ASGR-2. In some embodiments, the antigen binding proteins of the invention bind to pig ASGR-2. In some embodiments, the antigen binding proteins of the invention bind to dog ASGR-2. In some embodiments, the antigen binding proteins of the invention bind to mouse ASGR-2. In some embodiments, the antigen binding proteins of the invention bind to rat ASGR-2. In some embodiments, the antigen binding proteins specifically bind to ASGR-2 of different species.

[0099] In some embodiments, the antigen binding protein of the present invention binds to ASGR, ASGR-1 and / or ASGR-2 from two or more different species and / or binds to ASGR, ASGR-1 and / or ASGR-2 from the same species. For example, but not limited to, an antibody that binds to human and cynomolgus ASGR-1; an antibody that binds to human, cynomolgus and pig ASGR-1; an antibody that binds to human, cynomolgus, rat and mouse ASGR-2; an antibody that binds to human ASGR-1 and human ASGR-2; an antibody that binds to human and cynomolgus ASGR-1 and ASGR-2. In some embodiments, the antigen binding protein specifically binds to ASGR, ASGR-1 and / or ASGR-2 from two or more different species and / or specifically binds to ASGR, ASGR-1 and / or ASGR-2 from the same species.

[0100] As described herein, the ASGR receptor and ASGR-1 and / or ASGR-2 are separately internalized into cells after ligand binding. Thus, in certain embodiments, the present invention provides antigen binding proteins that inhibit or reduce the internalization of ASGR, ASGR-1 and / or ASGR-2. In certain embodiments, the antigen binding proteins of the present invention reduce ligand binding and inhibit the internalization of ASGR, ASGR-1 and / or ASGR-2. In some embodiments, the antigen binding proteins of the present invention inhibit internalization without necessarily inhibiting ligand binding.

[0101] In some embodiments, the antigen binding proteins (e.g., antibodies) of the present invention are pH and / or calcium insensitive molecules and bind ASGR, ASGR-1 and / or ASGR-2 and inhibit binding to ligands. It is envisioned that these properties are desirable to reduce or prevent dissociation of the molecule from the receptor during the endocytosis process in order to extend the half-life of the molecule. In some embodiments, the antigen binding proteins (e.g., antibodies) exhibit pH-independent binding to their antigen such that their affinity for antigen binding at physiological pH (i.e., pH 7.4) is similar to that at endosomal pH (i.e., pH 5.5-6.0). In some embodiments, the antigen binding proteins (e.g., antibodies) exhibit calcium-independent binding to their antigen such that their affinity for antigen binding at assay conditions (i.e., 1 mM calcium) is similar to that in the absence of exogenously added calcium. In some embodiments, the antigen binding protein exhibits both pH and calcium independent binding to its antigen, such that the affinity for antigen binding in the presence of calcium at physiological pH is similar to that in the absence of calcium at endosomal pH (i.e., pH 5.5-6.0). Any method known to one of skill in the art may be used to measure pH and / or calcium insensitivity, such as the method described in Example 7C below.

[0102] ASGR-1, an asialoglycoprotein receptor, contains an N-terminal cytosolic domain, a transmembrane domain, a stalk region, and a carbohydrate recognition domain (CRD) (alternatively known as the carbohydrate binding domain or "CBD"). The carbohydrate recognition domain ("CRD") structure of ASGR-1 has been reported in the literature (M. Meier et al., JMB (2000) 300, 857-865). Provided herein are structures of ASGR-1 at higher resolution than reported and when bound to various ligands (e.g., sugars, including but not limited to lactose, galactose, and / or GalNAc, or glycoproteins that display such sugars, including but not limited to fetuin, orosomucoid, and / or alkaline phosphatase) (see Example 10 and Figures 18-21 herein). Given the importance of this domain to the function of ASGR-1, in some embodiments it is desirable to target this domain with the antigen binding proteins of the invention.

[0103] Thus, in some embodiments, the antigen binding proteins of the invention bind to the CBD of ASGR-1. In certain embodiments, the antigen binding proteins of the invention bind to the CBD of human ASGR-1. In certain embodiments, the antigen binding proteins of the invention bind to the CBD of SEQ ID NO:5. In some embodiments, the antigen binding proteins of the invention bind to amino acid residues selected from the group consisting of 148-291, 149-291, 150-291, 151-291, 152-291, 153-291 and 154-291 of SEQ ID NO:5. In some embodiments, the invention comprises an isolated antigen binding protein that binds to the human ASGR-1 CBD within helix alpha-1 or helix alpha-2. In some embodiments, the invention comprises an isolated antigen binding protein that binds to the human ASGR-1 CBD within residues 174-186 of SEQ ID NO:5. In some embodiments, the invention includes isolated antigen binding proteins that bind to the human ASGR-1 CBD within residues 194-206 of SEQ ID NO:5. In some embodiments, the invention includes isolated antigen binding proteins that bind to the human ASGR-1 CBD at the same or overlapping binding site as that where a ligand (e.g., a sugar, including but not limited to, lactose, galactose, and / or GalNAc, or a glycoprotein that presents such a sugar, including but not limited to, fetuin, orosomucoid, and / or alkaline phosphatase, or other sugars and glycoproteins that can bind to ASGR, ASGR-1, and / or ASGR-2) binds. In some embodiments, the invention includes isolated antigen binding proteins that bind to the human ASGR-1 CBD within residues 237-273 or residues 240-267 of SEQ ID NO:5. In some embodiments, the antigen binding proteins of the invention bind to the CBD of cynomolgus monkey ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to the CBD of porcine ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to the CBD of canine ASGR-1. In some embodiments, the antigen binding proteins of the invention bind to the CBD of mouse ASGR-1.In some further embodiments, the antigen binding proteins of the invention bind to the CBD of rat ASGR-1. In some further embodiments, the antigen binding proteins of the invention bind to the CBD of two or more different ASGR-1 species, including, but not limited to, human ASGR-1 and cynomolgus monkey ASGR-1, or human ASGR-1, cynomolgus monkey ASGR-1 and dog ASGR-1, or human ASGR-1 and mouse ASGR-1.

[0104] In further embodiments, the antigen binding proteins of the invention bind to ASGR-1 and inhibit binding of ligands to ASGR-1. In specific embodiments, the ligands that are inhibited include, but are not limited to, sugars, including but not limited to, lactose, galactose and / or GalNAc, or glycoproteins that present such sugars, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase, or other sugars and glycoproteins that can bind to ASGR, ASGR-1 and / or ASGR-2.

[0105] The tyrosine at position 272 of mouse ASGR-1 (position 273 of human ASGR-1 (SEQ ID NO:5)) appears to be important for protein stability as it displays a hydrogen bond to D266 of mouse ASGR-1 and several van der Waals contacts to other residues of mouse ASGR-1 (N208, W210, H256 and R270). Moreover, by analogy with other lectins, Y272 of mouse ASGR-1 may play a role in carbohydrate binding and function of ASGR-1. Thus, in some embodiments, the antigen binding proteins of the present invention bind to or interact with Y273 of human ASGR-1. In some embodiments, the antigen binding proteins of the present invention bind to ASGR-1 at an epitope that includes Y273 of human ASGR-1. In some embodiments, the antigen binding proteins of the present invention bind to ASGR-1 at an epitope that renders Y273 of human ASGR-1 unable to participate in ligand binding.

[0106] Analysis of the crystal structure of hASGR-1 revealed specific amino acids involved in the interaction between hASGR-1 and ligands (e.g., sugars, including but not limited to, lactose, galactose, and / or GalNAc, or glycoproteins that present such sugars, including but not limited to, fetuin, orosomucoid, and / or alkaline phosphatase).Thus, in further embodiments, the antigen-binding proteins of the invention bind to or interact with at least one of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least one of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein that displays such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least one of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264.

[0107] In further embodiments, the antigen binding proteins of the invention bind to or interact with at least one of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least one of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein that displays such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least one of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.

[0108] In some embodiments, antigen binding proteins of the invention bind to or interact with at least two of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least three of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least four of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least five of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least six of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.In some embodiments, antigen binding proteins of the invention bind to or interact with at least seven of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least eight of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, an antigen binding protein of the invention binds to or interacts with at least nine of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least ten of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least eleven of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.In some embodiments, antigen binding proteins of the invention bind to or interact with at least twelve of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least 13 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least 14 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least 15 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least 16 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.In some embodiments, antigen binding proteins of the invention bind to or interact with at least 17 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least 18 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least 19 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, antigen binding proteins of the invention bind to or interact with at least twenty of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.

[0109] In some embodiments, the antigen binding proteins of the invention bind to or interact with at least two of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind to or interact with at least three of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind to or interact with at least four of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind to or interact with at least five of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind to or interact with at least six of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind to or interact with at least seven of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, antigen binding proteins of the invention bind to or interact with at least eight of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, antigen binding proteins of the invention bind to or interact with at least nine of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.In some embodiments, the antigen binding proteins of the invention bind to or interact with at least 10 of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind to or interact with at least 11 of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind to or interact with at least all of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.

[0110] In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least two of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least three of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least four of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least five of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least six of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least seven of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least eight of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least nine of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least ten of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least eleven of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 12 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 13 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 14 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 15 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 16 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 17 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 18 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least 19 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least twenty of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263, or W264.

[0111] In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least two of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least three of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least four of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least five of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least six of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least seven of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least eight of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the invention bind at an epitope that includes at least nine of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least 10 of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes at least 11 of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In some embodiments, the antigen binding proteins of the present invention bind at an epitope that includes all of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.

[0112] In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein presenting such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least two of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein that displays such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least three of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein that displays such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least four of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264.In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein that displays such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least five of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein that displays such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least six of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a saccharide including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein presenting such a saccharide including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least seven of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264.In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., a sugar, including but not limited to, lactose, galactose and / or GalNAc, or a glycoprotein that displays such a sugar, including but not limited to, fetuin, orosomucoid and / or alkaline phosphatase) with at least eight of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least nine of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least ten of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264.In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 11 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least twelve of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 13 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 14 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264.In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 15 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 16 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 17 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 18 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264.In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction between a ligand (e.g., lactose, galactose and / or GalNAc) and at least 19 of Q240, D242, W244, E253, N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction between a ligand (e.g., lactose, galactose and / or GalNAc) and at least 19 of Q240, D242, W244, E25. 3, blocks or reduces binding or interaction with at least 20 of N265, D266, D267, N209, R237, P238, E239, P241, D243, Y245, G246, H247, G252, C255, H257, T259, D260, V268, R271, Y273, F258, R263 or W264.

[0113] In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least two of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least three of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least four of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least five of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In a further embodiment, the antigen binding protein of the invention binds to hASGR-1 and blocks or reduces binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least six of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least seven of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least eight of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least nine of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In further embodiments, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least ten of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259 or Y273. In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with at least 11 of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.In a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with all of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273.

[0114] To correlate unique antigen-binding protein sequence features with specific functions or binding characteristics, sequences from the antigen-binding proteins of the present invention from various characterization bins may be analyzed. For example, the antigen-binding proteins of the present invention may be examined for their ability to bind to various binning probes (e.g., membrane preparations or soluble huASGR-1 from cells expressing ASGR-1 from different species). For each unique binding bin, the heavy and light chain sequences from each of the antigen-binding proteins may be compared and cladized based on, for example: 1. unique VDJ and VJ rearrangements; 2. divergence from germline (i.e., unique somatic hypermutations); and 3. relatedness to other antigen-binding proteins in the same bin. Thus, in certain embodiments, antigen-binding proteins that contain the same or similar sequence features and patterns will have substantially the same or similar binding characteristics. In specific embodiments, these antigen-binding proteins may bind to the same or similar epitopes with varying affinities.

[0115] Exemplary antigen binding proteins described herein have properties based on the epitopes in ASGR, ASGR-1 and / or ASGR-2 that are bound by the antigen binding protein. The term "epitope" includes any determinant that can be bound by an antigen binding protein, such as an antibody. An epitope is a region of an antigen that is bound by or interacts with an antigen binding protein that targets the antigen, and when the antigen is a protein, an epitope includes specific amino acids that directly contact or interact with the antigen binding protein. Epitopes can be formed both by contiguous amino acids or non-contiguous amino acids juxtaposed by tertiary folding of a protein. A "linear epitope" is an epitope in which the primary amino acid sequence comprises the epitope that is recognized. A linear epitope typically includes at least 3 or at least 4, more usually at least 5 or at least 6 or at least 7, e.g., about 8 to about 10, amino acids in a unique sequence.

[0116] A "conformational epitope", in contrast to a linear epitope, is a group of discontinuous amino acids (e.g., amino acid residues in a polypeptide that are not adjacent in the primary sequence of the polypeptide but are sufficiently close to each other in the context of the polypeptide's tertiary and quaternary structure to be bound by an antigen-binding protein). Epitopic determinants can include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and can have specific three-dimensional structural features and / or specific charge characteristics. In general, an antigen-binding protein specific for a particular target molecule preferentially recognizes an epitope on the target molecule in a complex mixture of proteins and / or macromolecules.

[0117] Methods for characterizing the epitope bound by an antigen binding protein are well known in the art and include, for example, binning (competition and / or cross-competition) (Miller et al., "Epitope binning of murine monoclonal antibodies by a multiplexed pairing assay," J Immunol Methods (2011) 365, 118-25), peptide mapping (e.g., PEPSPOT™) (Albert et al., "The B-cell Epitope of the Monoclonal Anti-Factor VIII Antibody ESH8 Characterized by Peptide Array Analysis," 2008 Thromb Haemost 99, 634-7), mutagenesis methods such as chimeras (Song et al., "Epitope Mapping of Ibalizumab, a Humanized Anti-CD4 Monoclonal Antibody with Anti-HIV-1 Activity in Infected Patients," J. Virol.(2010) 84, 6935-6942), alanine scanning (Cunningham and Wells, "High-resolution epitope mapping of HGH-receptor interactions by alanine-scanning mutagenesis," Science (1989) 244, 1081-1085), arginine scanning (Lim et al., "A diversity of antibody epitopes can induce signaling through the erythropoietin receptor," Biochemistry (2010) 49, 3797-3804), HD exchange method (Coates et al., "Epitope mapping by amide hydrogen / deuterium exchange coupled with immobilization of antibody, on-line proteolysis, liquid chromatography and mass spectrometry," Rapid Commun. Mass Spectrom. (2009) 23, 639-647), NMR cross-saturation method (Morgan et al., "Precise epitope mapping of malaria parasite inhibitory antibodies" These methods include, but are not limited to, crystallography (Gerhardt et al. "Structure of IL-17A in complex with a potent, fully human neutralizing antibody" J. Mol. Biol (2009) 394, 905-21). These methods vary in the level of detail they provide regarding the amino acids that comprise the epitope.

[0118] Antigen binding proteins of the present invention include those that have the same or overlapping epitopes as the exemplary antigen binding proteins listed in Tables 2-7. In some embodiments, the antigen binding proteins have the same epitopes as the exemplary antigen binding proteins. In other embodiments, the antigen binding proteins bind only to the same subset of amino acids as the exemplary antigen binding proteins. In some embodiments, antigen binding proteins that can bind to any of the epitopes bound by the antibodies listed in Tables A, B, C, or 6 are particularly useful.

[0119] In certain embodiments, an antigen binding protein of the present invention has an epitope identical or overlapping with an antigen binding protein in Table 2-7 and comprises: a) a light chain variable domain that has at least 90% identity, at least 95% identity or identical to the amino acid sequence of an antigen binding protein set forth in Table 2-7; b) a heavy chain variable domain that has at least 90% identity, at least 95% identity or identical to the amino acid sequence of an antigen binding protein set forth in Table 2-7; or c) the light chain variable domain of a) and the heavy chain variable domain of b).

[0120] In certain embodiments, the antigen binding proteins of the invention have an epitope identical or overlapping with an antigen binding protein selected from the group consisting of 25A4, 4H6, 4A2, 5E5, 7E11, 54E9, 22G5, 194A4, 218G4, 176H4 and 194C10, and the antigen binding protein has a light chain variable domain that is at least 90% identical, at least 95% identical or identical to the amino acid sequence of 25A4, and a light chain variable domain that is at least 90% identical to the amino acid sequence of 25A4. a heavy chain variable domain that is at least 90% identical, at least 95% identical, to the amino acid sequence of 4H6; a light chain variable domain that is at least 90% identical, at least 95% identical, to the amino acid sequence of 4H6; and a heavy chain variable domain that is at least 90% identical, at least 95% identical, to the amino acid sequence of 4H6; a light chain variable domain that is at least 90% identical, at least 95% identical, to the amino acid sequence of 4A2. a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 5E5, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 5E5; a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 7E11, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 7E11; a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 54E9, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 54E9;a light chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 22G5, and a heavy chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 22G5; a light chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 194A4, and a heavy chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 194A4; a light chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 218G4G4, and a heavy chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 18G4; a light chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 176H4, and a heavy chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 176H4; a light chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 194C10, and a heavy chain variable domain having at least 90% identity, at least 95% identity or identical to the amino acid sequence of 194C10;

[0121] In certain embodiments, the ASGR-1 antigen binding proteins of the invention have an epitope identical or overlapping with an antibody in Tables 2-7 and comprise a light chain variable domain comprising: a) an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in Table 2; a LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in Table 2; and a LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in Table 2; and a heavy chain variable domain comprising: a) an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in Table 2; a HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in Table 2; and a HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in Table 2.

[0122] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with an antibody in Table A, B, C or 6 and comprises a light chain variable domain comprising: a) an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in Table A, B, C or 6; a LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in Table A, B, C or 6; and a LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in Table A, B, C or 6; and a heavy chain variable domain comprising: a) an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in Table A, B, C or 6; a HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in Table A, B, C or 6; and a HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in Table A, B, C or 6.

[0123] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 25A4 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:480; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8492; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:16504; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4488; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12500; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:20512.

[0124] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 4H6 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:894; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8906; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:16918; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4902; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12914; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:20926.

[0125] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 4A2 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:1130; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:9142; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:17154; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:5136; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:13148; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:21160.

[0126] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 5E5 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:974; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8986; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:16998; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4982; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12994; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:21006.

[0127] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with antibody 7E11 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:872; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8884; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:16896; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4880; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12892; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:20904.

[0128] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 54E9 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:3448; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:11460; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:19472; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:7452; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:15464; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:23476.

[0129] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 22G5 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:326; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8338; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:16350; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4334; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12346; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:20358.

[0130] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 194A4 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:2780; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:10792; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:18804; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:6786; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:14798; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:22810.

[0131] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 218G4 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:3746; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:11758; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:19770; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:7750; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:15762; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:23774.

[0132] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with that of antibody 176H4 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:2502; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:10514; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:18526; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:6508; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:14520; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:22532.

[0133] In certain embodiments, an ASGR-1 antigen binding protein of the invention has an epitope identical or overlapping with antibody 194C10 and comprises a light chain variable domain comprising an LCDR1 having no more than 3 amino acid additions, deletions or substitutions from the LCDR1 sequence set forth in SEQ ID NO:2792; an LCDR2 having no more than 3 amino acid additions, deletions or substitutions from the LCDR2 sequence set forth in SEQ ID NO:10804; and an LCDR3 having no more than 3 amino acid additions, deletions or substitutions from the LCDR3 sequence set forth in SEQ ID NO:18816; and a heavy chain variable domain comprising an HCDR1 having no more than 3 amino acid additions, deletions or substitutions from the HCDR1 sequence set forth in SEQ ID NO:6798; an HCDR2 having no more than 3 amino acid additions, deletions or substitutions from the HCDR2 sequence set forth in SEQ ID NO:14810; and an HCDR3 having no more than 3 amino acid additions, deletions or substitutions from the HCDR3 sequence set forth in SEQ ID NO:22822.

[0134] Antigen binding proteins with identical or overlapping epitopes often compete with each other for binding to the antigens ASGR, ASGR-1 and / or ASGR-2. Thus, in certain embodiments, the antigen binding proteins of the invention (e.g., antibodies or antibody fragments thereof) compete with the antigen binding proteins listed in Tables 2-7. In some embodiments, the antigen binding proteins of the invention (e.g., antibodies or antibody fragments thereof) compete with the antigen binding proteins listed in Tables A, B and C. In some embodiments, the antigen binding proteins of the invention (e.g., antibodies or antibody fragments thereof) compete with the antigen binding proteins listed in Table 6. "Compete" or "competition" means that the antigen binding proteins compete for the same epitope or binding site on a target. Such competition can be determined by an assay in which a reference antigen binding protein (e.g., an antibody or antibody fragment thereof) prevents or inhibits specific binding of the test antigen binding protein. Many types of competitive binding assays may be used to determine whether a test molecule competes with a reference molecule for binding.Examples of assays that can be used include solid-phase direct or indirect radioimmunoassays (RIA), solid-phase direct or indirect enzyme immunoassays (EIA), sandwich competitive assays (see, e.g., Stahli et al. (1983) Methods in Enzymology 9:242-253), solid-phase direct biotin-avidin EIA (see, e.g., Kirkland et al. (1986) J. Immunol. 137:3614-3619), solid-phase direct labeling assays, solid-phase direct labeling sandwich assays, Luminex (Jia et al., "A novel method of Multiplexed Competitive Antibody Binning for the characterization of monoclonal antibodies," J. Immunological Methods (2004) 288, 91-98), and surface plasmon resonance (Song et al., "Epitope Mapping of Ibalizumab, a Humanized Anti-CD4 Monoclonal Antibody with Anti-HIV-1 Activity in Infected ("Competition in Patients" J. Virol. (2010) 84, pp. 6935-6942). Exemplary methods for determining competition are described in Example 7D. Typically, when a competing antigen-binding protein is present in excess, it inhibits binding of the reference antigen-binding protein to the common antigen by at least 50%, 55%, 60%, 65%, 70% or 75%. In some cases, binding to ASGR-1 is inhibited by at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more.

[0135] In addition to competition, antigen binding proteins (e.g., antibodies or antibody fragments thereof) with identical, overlapping, or similar epitopes may be similarly affected by mutagenesis of ASGR, ASGR-1, and / or ASGR-2. Briefly, the domain(s) / region(s) containing residues that contact or are buried in the antibody can be identified by mutating specific residues in ASGR, ASGR-1, and / or ASGR-2 (e.g., wild-type antigen) and determining whether the antigen binding protein can bind to the mutated or variant ASGR, ASGR-1, and / or ASGR-2 protein. By making a large number of individual mutations, residues that play a direct role in binding or are in close enough proximity to the antibody that the mutations can affect binding between the antigen binding protein and the antigen can be identified. Knowledge of these amino acids can reveal the domain(s) or region(s) of the antigen that contact the antigen binding protein or contain residues that fit the antibody. Such domains can contain the binding epitopes of the antigen binding protein. As mentioned above, one specific example of this general approach utilizes an arginine / glutamic acid scanning protocol (see, e.g., Naneviz, T. et al., 1995, J.Biol.Chem., 270:37, pp. 21619-21625; and Zupnick, A. et al., 2006, J.Biol.Chem., 281:29, pp. 20464-20473). In general, arginine and glutamic acid are substituted (typically individually) for amino acids in the wild-type polypeptide because these amino acids are charged and bulky, and therefore have the potential to disrupt binding between the antigen-binding protein and the antigen in the region of the antigen where the mutation is introduced. Arginine residues present in the wild-type antigen are replaced with glutamic acid. A variety of such individual mutants are obtained and the collected binding results are analyzed to determine which residues affect binding.In Example 7E, arginine / glutamic acid mutagenesis scanning was performed using the human ASGR-1 CBD domain to determine the effect on the exemplary antibody. ASGR, ASGR-1 and / or ASGR-2 antigen binding proteins with characteristics that are affected in a similar manner to the exemplary antibody by mutagenesis are included within the scope of the invention.

[0136] Example 7E describes one example of such arginine / glutamic acid scanning of ASGR-1 against the ASGR-1 antigen binding proteins provided herein. A series of mutant ASGR-1 antigens were generated, each mutant antigen having a single mutation. Binding of each mutant ASGR-1 antigen by various ASGR-1 antigen binding proteins was measured and compared to the ability of the selected antigen binding proteins to bind to human ASGR-1 (SEQ ID NO:5). In certain embodiments, binding of the antigen binding proteins of the invention to ASGR-1 is inhibited by a single mutation in ASGR-1, the single mutation being selected from the set of SEQ ID NO:5. As shown in NO:5, it is selected from the group consisting of R170, S171, G172, R183, L184, W195, E196, K199, H203, H204, P207, V208, N209, H215, D216, P220, D225, D228, R237, P238, E239, P241, D242, D243, Y245, G246, H247, G248, L249, G251, E253, T259, D260, R263, N265, Q270, R271, P272, R274, and E280. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 4A2, and its binding to ASGR-1 is inhibited by any of the following mutations: W195, E196, K199, H204, P207, and R263. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 4B3, and its binding to ASGR-1 is inhibited by any of the following mutations: H203, H204, P220, and G251. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 5E5, and its binding to ASGR-1 is inhibited by any of the following mutations: W195, K199, and R263. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 6G7, whose binding to ASGR-1 is inhibited by any of the following mutations: R183, L184, H215, P220, P238, G246, H247, G248, G251, and N265.In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 149D11, and its binding is inhibited by any of the mutations at R170, S171, and L184. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 175F4, and its binding is inhibited by any of the mutations at R183. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 17H6, and its binding is inhibited by any of the mutations at P241, D242, D243, Y245, G251, and E253. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 194A4, and its binding is inhibited by any of the mutations at D260. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 60C12, whose binding is inhibited by any of the following mutations: R170, R237, E239, P241, T259, D260, R263, and N265. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 65D5, whose binding is inhibited by any of the following mutations: R237, T259, D260, and R263. In some embodiments, the ASGR-1 antigen binding protein shares the attributes of antibody 190F8 or 191G1, and its binding is inhibited by any of the following mutations: R170, S171, G172, E196, H204, P207, V208, N209, H215, D216, D225, D228, P238, D243, G248, L249, G251, D260, Q270, R271, P272, R274, and E280. In some embodiments, the ASGR-1 antigen binding protein shares the attributes of antibody 199A7, and its binding is inhibited by any of the following mutations: R170, R183, H215, and Q270. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 146B6, whose binding is inhibited by any of the mutations at P241, T259, and N265. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 193E7, whose binding is inhibited by any of the mutations at P207 and R263.In some embodiments, any of 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 of the single mutations in the above groups individually inhibit binding of the ASGR-1 antigen binding protein to ASGR-1.

[0137] The binding of various anti-ASGR-1 antigen binding proteins (e.g., antibodies 5E5, 22G5, 7E11, 4A2, 4H6, 72G9, 194A4, 54E9, 218G4, 176H4 and 194C10) was further analyzed using X-ray crystallography. The X-ray crystallography results were highly correlated with the arginine / glutamic acid mutagenesis profiling results described above and in Example 7E. The interface between an antigen binding protein and an antigen can be determined / defined in a number of ways. In Examples 10B-L, the interface was determined by selecting interface residues that have at least one atom within a predefined distance with their partner protein. In some embodiments, ASGR-1 residues present within the interface with antibody 5E5, as determined by a distance of 8 Å or less, are H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, V159, E160, R163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260, or W264 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 5E5, as determined by a distance of 5 Å or less, are H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262 or R263 (SEQ ID NO:5).In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 5E5, including any of H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237, P238, D261, G262, R263, V159, E160, R163, T193, S194, E197, V201, I205, G206, P207, Y229, E230, T231, E239, F258, T259, D260, or W264 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 5E5, including any of H161, E162, W195, E196, Q198, K199, F200, Q202, H203, H204, G232, F233, K234, N235, W236, R237 or P238 (SEQ ID NO:5) present within the interface.

[0138] In some embodiments, the ASGR-1 residues present within the interface with antibody 22G5, as determined by a distance of 8 Å or less, are W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, P155, N157, W158, F168, S169, R170, W175, A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, or T279 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 22G5, as determined by a distance of 5 Å or less, are W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272 or W275 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 5E5, including any of W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272, W275, P155, N157, W158, F168, S169, R170, W175, A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, or T279 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 5E5, including any of W167, S171, G172, K173, A174, A176, D177, N180, Y181, R183, L184, E185, D186, Q270, P272 or W275 (SEQ ID NO:5) present within the interface.

[0139] In some embodiments, the ASGR-1 residues present within the interface with antibody 4A2, as determined by a distance of 8 Å or less, are R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, P155, N157, W158, F168, S169, R170, W175, A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, or T279 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 4A2, as determined by a distance of 5 Å or less, are R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263 or R274 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 4A2, including any of R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274, P155, N157, W158, F168, S169, R170, W175, A178, D179, C182, A187, W211, C269, R271, Y273, R274, C277, or T279 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 4A2, including any of R170, W195, E196, K199, Q202, H203, H204, I205, G206, P207, V208, F233, K234, N235, W236, P238, D260, D261, G262, R263, R274 (SEQ ID NO:5) present within the interface.

[0140] In some embodiments, ASGR-1 residues present within the interface with antibody 7E11, as determined by a distance of 8 Å or less, are H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204, Y229, E230, T231, G232, E239, Q240, P241, D261, G262, or W264 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 7E11, as determined by a distance of 5 Å or less, are H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238 or R263 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 7E11, including any of H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238, R263, E160, E162, V192, T193, E197, V201, H204, Y229, E230, T231, G232, E239, Q240, P241, D261, G262, or W264 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 7E11, including any of H161, S194, W195, E196, Q198, K199, F200, Q202, H203, F233, K234, N235, W236, R237, P238 or R263 (SEQ ID NO:5) present within the interface.

[0141] In some embodiments, ASGR-1 residues present within the interface with antibody 4H6, as determined by a distance of 8 Å or less, are H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260, G262, or W264 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 4H6, as determined by a distance of 5 Å or less, are H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261 or R263 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 4H6, including any of H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261, R263, R163, V192, E197, Q198, H203, P207, D228, E230, W236, R237, D260, G262, or W264 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 4H6, including any of H161, E162, T193, S194, W195, E196, K199, Q202, T231, G232, F233, K234, N235, P238, D261 or R263 (SEQ ID NO:5) present within the interface.

[0142] In some embodiments, ASGR-1 residues present within the interface with antibody 72G9, as determined by a distance of 8 Å or less, are D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268, or C269 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 72G9, as determined by a distance of 5 Å or less, are D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, or Q270 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 72G9, including any of D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, Q270, H215, K222, T231, G232, R237, P238, H247, G248, E253, C255, D266, V268, C269 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 72G9, including any of D216, Q217, N218, G219, P220, W221, Y229, E230, K234, W236, E239, Q240, P241, D242, D243, W244, Y245, G246, L249, G250, G251, G252, D254, or Q270 (SEQ ID NO:5) present within the interface.

[0143] In some embodiments, the ASGR-1 residues present within the interface with antibody 194A4, as determined by a distance of 8 Å or less, are T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, H161, E162, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263, or W264 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 194A4, as determined by a distance of 5 Å or less, are T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, or G252 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 194A4, including any of T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, G252, H161, E162, V191, V192, E197, Q198, D216, G219, K222, W223, D225, R263, or W264 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 194A4, including any of T193, S194, W195, E196, P220, W221, G226, T227, D228, Y229, E230, T231, G232, F233, K234, N235, W236, R237, P238, E239, or G252 (SEQ ID NO:5) present within the interface.

[0144] In some embodiments, the ASGR-1 residues present within the interface with antibody 194C10, as determined by a distance of 8 Å or less, are N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 194C10, as determined by a distance of 5 Å or less, are N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273 or R274 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 194C10 and is selected from the group consisting of N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, R274, V156, W158, V159, H161, W167, F168, S169, K173, K199, F200, V201, W211, R237, H257, F258, T259, D261, D267, V268, Q270, or W275 (SEQ ID NO: 1). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 194C10, including those in which any of N157, R170, S171, G172, Q202, H203, H204, I205, G206, P207, V208, N209, T210, D260, R271, P272, Y273, or R274 (SEQ ID NO:5) are present within the interface.

[0145] In some embodiments, ASGR-1 residues present within the interface with antibody 54E9, as determined by a distance of 8 Å or less, are W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264, or D266 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 54E9, as determined by a distance of 5 Å or less, are W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271 or Y273 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 54E9, including any of W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, Y273, Q198, Q202, P207, V208, F233, W236, D243, E253, F258, G262, W264, or D266 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 54E9, including any of W195, N209, N235, R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271 or Y273 (SEQ ID NO:5) present within the interface.

[0146] In some embodiments, ASGR-1 residues present within the interface with antibody 218G4, as determined by a distance of 8 Å or less, are R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269, or W275 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 218G4, as determined by a distance of 5 Å or less, are R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273 or R274 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 218G4, including any of R170, S171, G172, A174, H204, I205, G206, P207, V208, N209, H257, D260, N265, D267, Q270, R271, P272, Y273, R274, W167, F168, S169, K173, W175, D177, Y181, Q202, H203, T210, W211, R237, F258, T259, D261, D266, V268, C269, or W275 (SEQ ID NO:5) present within the interface. In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 218G4, including any of R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273 or R274 (SEQ ID NO:5) present within the interface.

[0147] In some embodiments, the ASGR-1 residues present within the interface with antibody 176H4, as determined by a distance of 8 Å or less, are R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D26 0, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, or W275 (SEQ ID NO:5). In some embodiments, ASGR-1 residues present within the interface with antibody 176H4, as determined by a distance of 5 Å or less, are R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273 or R274 (SEQ ID NO:5). In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 176H4 and includes R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T2 59, D260, N265, D267, Q270, R271, P272, Y273, R274, S169, W175, A176, A178, T210, W211, W236, P238, E239, D242, Y245, G250, G251, F258, D261, G262, R263, W264, D266, V268, C269, W275 (SEQ ID NO:5) are present within the interface.In certain embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 176H4, including any of R170, S171, G172, K173, A174, D177, P207, V208, N209, R237, Q240, W244, G246, H247, G248, L249, E253, H257, T259, D260, N265, D267, Q270, R271, P272, Y273, R274 (SEQ ID NO:5) present within the interface.

[0148] In some embodiments, ASGR-1 residues involved in ligand binding are also in close proximity to the area where antibodies 72G9, 54E9, 218G4 or 176H4 bind, and may be useful for engineering ASGR-1 binding to ligands. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 72G9 and a ligand (e.g., GalNAc), including any of Q240, D242, W244, E239, P241, D243, Y245, G246, G252, R237, E253, P238, H247, C255 or V268 (SEQ ID NO:5) present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 72G9 and a ligand (e.g., GalNAc), including one in which any of Q240, D242, or W244 (SEQ ID NO:5) are present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 72G9 and a ligand (e.g., GalNAc), including one in which any of Q240, D242, W244, E239, P241, D243, Y245, G246, or G252 (SEQ ID NO:5) are present within the interface. In some embodiments, ASGR-1 antigen binding protein forms an interface with ASGR-1, which overlaps with the interface of antibody 72G9 and ligand (e.g., GalNAc), including any of Q240, D242, W244, R237 or E253 (SEQ ID NO:5) being present within the interface. As described in the following examples, the degree of inhibition caused by 72G9 is lower than other direct blocking antibodies provided herein. Without intending to be limiting, it is understood that this occurs due to the nature of the relative orientation of ASGR-1 protein and antibody when bound to each other. For example, when 72G9 antibody is bound to ASGR-1, there is still enough space for the ligand to reach the binding site to some extent (albeit less).In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 54E9 and a ligand (e.g., GalNAc), including any of N209, R237, Q240, D242, H257, T259, N265, D267, Y273, P238, E239, D260, R263, R271, E253, D266, D243, F258 or W264 (SEQ ID NO:5) present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 54E9 and a ligand (e.g., GalNAc), including any of N209, R237, Q240, D242, H257, T259, N265, D267, or Y273 (SEQ ID NO:5) present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 54E9 and a ligand (e.g., GalNAc), including any of N209, R237, Q240, D242, H257, T259, N265, D267, Y273, P238, E239, D260, R263, or R271 (SEQ ID NO:5) present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 54E9 and a ligand (e.g., GalNAc), including one in which any of N209, R237, Q240, D242, H257, T259, N265, D267, Y273, E253, or D266 (SEQ ID NO:5) are present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 218G4 and a ligand (e.g., GalNAc), including one in which any of N209, H257, N265, D267, Y273, D260, R271, R237, T259, D266, F258, or V268 (SEQ ID NO:5) are present within the interface.In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 218G4 and a ligand (e.g., GalNAc), including one in which any of N209, H257, N265, D267, or Y273 (SEQ ID NO:5) are present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 218G4 and a ligand (e.g., GalNAc), including one in which any of N209, H257, N265, D267, Y273, D260, or R271 (SEQ ID NO:5) are present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 218G4 and a ligand (e.g., GalNAc), including any of N209, H257, N265, D267, Y273, R237, T259 or D266 (SEQ ID NO:5) present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 176H4 and a ligand (e.g., GalNAc), including any of N209, R237, Q240, W244, E253, H257, T259, N265, D267, Y273, G246, H247, D260, R271, D266, P238, E239, Y245, F258, R263, W264 or V268 (SEQ ID NO:5) present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 176H4 and a ligand (e.g., GalNAc), including any of N209, R237, Q240, W244, E253, H257, T259, N265, D267 or Y273 (SEQ ID NO:5) present within the interface.In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 176H4 and a ligand (e.g., GalNAc), including those in which any of N209, R237, Q240, W244, E253, H257, T259, N265, D267, Y273, G246, H247, D260, or R271 (SEQ ID NO:5) are present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface of antibody 176H4 and a ligand (e.g., GalNAc), including those in which any of N209, R237, Q240, W244, E253, H257, T259, N265, D267, Y273, or D266 (SEQ ID NO:5) are present within the interface.

[0149] As described above, binding interactions between huASGR-1 and ligands (e.g., lactose, galactose, GalNAc), as well as between huASGR-1 and various embodiments of antigen binding proteins (e.g., antibodies) of the invention, were assessed using X-ray crystallog...

Claims

1. An isolated monoclonal antibody that binds to human ASGR-1 (asialoglycoprotein receptor 1), comprising the amino acid sequence of SEQ ID NO:5, wherein the isolated monoclonal antibody inhibits ASGR-1 binding to a ligand and binds to ASGR-1 with an equilibrium dissociation constant of 10 -8 M or less.

2. An isolated monoclonal antibody that binds to human ASGR-1 (asialoglycoprotein receptor 1) having the amino acid sequence of SEQ ID NO:5, and inhibits ASGR-1 binding to a ligand with a half-maximal inhibitory concentration (IC50) of 90 nM or less.

3. An isolated monoclonal antibody described in claim 1 or 2, which binds to the carbohydrate recognition domain of human ASGR-1.

4. An isolated monoclonal antibody described in claim 1 or 2, which inhibits internalization of ASGR (asialoglycoprotein receptor).

5. An isolated monoclonal antibody described in any one of claims 1 to 4, which further binds to ASGR-2 (asialoglycoprotein receptor 2).

6. An isolated monoclonal antibody according to any one of claims 1 to 5, comprising a VH CDR1, a VH CDR2, and a VH CDR3 having amino acid sequences identical to each CDR or containing one, two, or three amino acid residue substitutions in each CDR compared to an antibody VH of any of the sequences set forth in Tables 3 to 7.

7. The isolated monoclonal antibody of claim 6, comprising a VL CDR1, a VL CDR2, and a VL CDR3 having amino acid sequences identical to each CDR or containing one, two, or three amino acid residue substitutions in each CDR compared to an antibody VL of any of the sequences set forth in Tables 3-7.

8. The isolated monoclonal antibody of claim 7, wherein the VH CDR and the VL CDR are a pair of corresponding VH CDR and VL CDR as set forth in Table 2.

9. An isolated monoclonal antibody described in claim 8, comprising a heavy chain variable domain having at least 90% identity to any of the VH domain amino acid sequences listed in Tables 3 to 7.

10. An isolated monoclonal antibody described in claim 8 or 9, which comprises a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences listed in Tables 3 to 7.

11. The isolated monoclonal antibody of claim 10, wherein the light chain variable domain and the heavy chain variable domain are a corresponding VL and VH pair as set forth in Tables 3 to 7.

12. An isolated monoclonal antibody that competes with an isolated monoclonal antibody described in any one of claims 1 to 11 for binding.

13. An isolated monoclonal antibody described in any one of claims 1 to 12, which is a chimeric antibody, a humanized antibody, or a human antibody.

14. A pharmaceutical composition comprising an isolated monoclonal antibody according to any one of claims 1 to 13 and a pharma- ceutical acceptable excipient.

15. An isolated nucleic acid encoding an isolated monoclonal antibody described in any one of claims 1 to 13.

16. A vector comprising the nucleic acid described in claim 15.

17. A host cell comprising the vector described in claim 16 or the nucleic acid described in claim 15.

18. A method for producing an antibody, comprising culturing a host cell described in claim 17 and recovering the antibody.

19. A pharmaceutical composition for reducing the risk of cardiovascular disease in a patient in need thereof, comprising a therapeutically effective dose of an isolated monoclonal antibody or antigen-binding fragment thereof described in any one of claims 1 to 13, wherein the isolated monoclonal antibody or antigen-binding fragment thereof binds to human ASGR-1, inhibits binding between human ASGR-1 and GalNAc residues of nonsialylated glycoproteins, and reduces LDL cholesterol levels.

20. The pharmaceutical composition of claim 19, wherein the cardiovascular disease is coronary artery disease or myocardial infarction.

21. A pharmaceutical composition for lowering LDL cholesterol levels in a patient in need thereof, comprising a therapeutically effective dose of an isolated monoclonal antibody or antigen-binding fragment thereof described in any one of claims 1 to 13.

22. A pharmaceutical composition for lowering non-HDL cholesterol levels in a patient in need thereof, comprising a therapeutically effective dose of an isolated monoclonal antibody or antigen-binding fragment thereof described in any one of claims 1 to 13.

23. A pharmaceutical composition for increasing serum ALP (alkaline phosphatase) levels in a patient, comprising a therapeutically effective dose of an isolated monoclonal antibody or antigen-binding fragment thereof described in any one of claims 1 to 13.

24. A pharmaceutical composition described in any one of claims 19 to 23, administered simultaneously or sequentially with at least one drug that reduces cholesterol.

25. The pharmaceutical composition of claim 24, wherein the at least one drug is a statin, an anti-PCSK9 inhibitor, or a combination thereof.

26. The pharmaceutical composition of claim 24 or 25, wherein the at least one drug is selected from the group consisting of evolocumab, alirocumab, bococizumab, ALN-PCS, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, simvastatin, and any combination thereof.

27. Use of an isolated monoclonal antibody or antigen-binding fragment thereof described in any one of claims 1 to 13 in the preparation of a medicament for treating or preventing cardiovascular disease in a patient in need of such treatment or prevention.