ASGR inhibitors

Abstract

This paper describes antigen-binding proteins that interact with ASGR, ASGR-1, and / or ASGR-2, as well as methods for producing and using such antigen-binding proteins. It also describes methods for treating and preventing cardiovascular disease by administering pharmaceutically effective amounts of ASGR, ASGR-1, and / or ASGR-2 antigen-binding proteins. Furthermore, it describes methods for treating and preventing cardiovascular disease by administering pharmaceutically effective amounts of interfering RNA compositions that reduce the expression of ASGR, ASGR-1, and / or ASGR-2.

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. Reference to electronic sequence listings and tables This application contains a Sequence Listing, which 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 related 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 heart or blood vessel, 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 cardiovascular system.Medicines for treating cardiovascular disease, especially coronary artery disease, have been introduced over the past few years (for example, 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 to human ASGR and inhibit ASGR function. In one embodiment, the present invention includes an isolated antigen-binding protein that binds to human ASGR and inhibits ASGR binding to a ligand. In another embodiment, the present invention includes an isolated antigen-binding protein that binds to human ASGR-1 and inhibits ASGR-1 binding to a ligand and / or ASGR-1 interaction with ASGR-2. In another embodiment, the present invention includes an isolated antigen-binding protein that binds to human ASGR-2 and inhibits ASGR-2 binding to a ligand and / or ASGR-2 interaction with ASGR-1. In yet another embodiment, the present invention includes an isolated antigen-binding protein that binds to human ASGR-1 and human ASGR-2 and inhibits ASGR-1 and / or ASGR-2 binding to a ligand. In some embodiments, the isolated binding protein specifically binds to human ASGR, ASGR-1, and / or ASGR-2.

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

[0006] In some aspects, the present invention provides isolated antigen binding proteins that specifically bind to 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 Tables 3-7. In some aspects, the present invention provides isolated antigen binding proteins that specifically bind to 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 Tables 3-7. In some embodiments, the present invention provides isolated antigen binding proteins that specifically bind to 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 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 isolated antigen binding proteins that specifically bind to human ASGR-1 and comprise 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 isolated antigen binding proteins that specifically bind to 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 A. In some embodiments, the invention provides isolated antigen binding proteins that specifically bind to 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 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 present 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 listed in Table 6, and a light chain variable domain having at least 90% identity to any of the VL domain amino acid sequences listed in Table 6. In some embodiments, the present 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 listed in Table 6, and a light chain variable domain having any of the VL domain amino acid sequences listed in Table 6.

[0007] In some aspects, the invention provides isolated antigen binding proteins that bind to human ASGR-1 and comprise one or more VH CDR1s, VH CDR2s, or VH CDR3s 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 isolated antigen binding proteins that bind to human ASGR-1 and comprise one or more VH CDR1s, VH CDR2s, or VH CDR3s having an amino acid sequence identical to, or containing conservative substitutions (substitutions) of, 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 to human ASGR-1 and comprises 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 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 to human ASGR-1 and comprises one or more VL CDR1, VL CDR2, or VL CDR3 having an amino acid sequence identical to, or containing conservative substitutions (substitutions) of, any of the amino acid sequences set forth in Table 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 of the VH 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 of the VL 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 conservative substitutions for, any of the amino acid sequences set forth in Table 19B or 19C, 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 conservative substitutions for, any of the amino acid sequences set forth in Table 20B or 20C, as depicted in Figure 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 conservative substitutions for, any of the amino acid sequences set forth in Tables 19B or 19C, as depicted in Figure 55, and one VL CDR1, VL CDR2, or VL CDR3 having an amino acid sequence identical to, or containing conservative substitutions for, 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 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 of the VHs 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 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 to human ASGR-1 and comprises two VH CDR1s, VH CDR2s, or VH CDR3s having amino acid sequences identical to, or containing conservative substitutions for, any of the amino acid sequences set forth in Tables 19B or 19C, as depicted in Figure 55, and two VL CDR1s, VL CDR2s, or VL CDR3s having amino acid sequences identical to, or containing conservative substitutions for, 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 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 of the VH 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 of the VL 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, VH CDR2, or VH CDR3 having an amino acid sequence identical to, or containing conservative substitutions for, any of the amino acid sequences set forth in Tables 19B or 19C, as depicted in Figure 55, and a VL CDR1, VL CDR2, or VL CDR3 having an amino acid sequence identical to, or containing conservative substitutions for, any of the amino acid sequences set forth in Tables 20B or 20C, as depicted in Figure 55.

[0008] In some aspects, the present invention provides isolated antigen binding proteins that specifically bind to 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 present invention provides isolated antigen binding proteins that specifically bind to 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 present invention provides antigen binding proteins that specifically bind to human ASGR-1 at an epitope bound by any of the antigen binding proteins disclosed herein. In some embodiments, the present 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 Tables 2-7. In some embodiments, the present 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 A. In some embodiments, the present 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 B. In some embodiments, the present 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 C. In some embodiments, the present invention provides isolated antigen binding proteins that specifically bind to human ASGR-1 at the epitope bound by at least one of the antigen binding proteins set forth in Table 6.

[0010] In some aspects, the present 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 present 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 present 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 present 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 present invention provides isolated antigen-binding proteins that compete for binding with any of the antigen-binding proteins set forth in Table C. In yet other embodiments, the present 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 present invention provides isolated antigen-binding proteins that bind to 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 to human ASGR-1 within residues 148-291, 149-291, 150-291, 151-291, 152-291, 153-291, 154-291, or 155-291 of SEQ ID NO:5. In some embodiments, the present invention includes isolated antigen-binding proteins that bind to human ASGR-1 CBD within helix alpha-1. In some embodiments, the present invention includes isolated antigen-binding proteins that bind to human ASGR-1 CBD within residues 174-186 of SEQ ID NO:5. In some embodiments, the present invention includes isolated antigen-binding proteins that bind to human ASGR-1 CBD within helix alpha-2. In some embodiments, the invention includes an isolated antigen binding protein that binds to human ASGR-1 CBD within residues 194-206 of SEQ ID NO:5. In some embodiments, the invention includes an isolated antigen binding protein that binds 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 that is 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 isolated antigen-binding proteins or antibodies that bind to human ASGR-1 and inhibit human ASGR-1 function. In some embodiments, the isolated antigen-binding proteins or antibodies bind to human ASGR-1 and inhibit human ASGR-1 from binding to a ligand. In some embodiments, the antigen-binding proteins or antibodies, or the paratopes in the antibodies, bind 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 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 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、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、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(SE, Q ID NO:5). In some embodiments, the antigen binding protein or antibody or 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 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,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 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 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 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, 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 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 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 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、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、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 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 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 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 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 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 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 the 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 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 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 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 the 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 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 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 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 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 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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 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 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 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 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 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 isolated antigen-binding proteins or antibodies or paratopes therein that specifically bind to human ASGR-1 and inhibit human ASGR-1 function. In some embodiments, the isolated antigen-binding proteins or antibodies or paratopes therein specifically bind to human ASGR-1 and inhibit human ASGR-1 from binding to a ligand. In some embodiments, the antigen-binding proteins or antibodies or paratopes therein specifically bind to human ASGR-1 within residues 148-291 of SEQ ID NO:5. In some embodiments, when the antigen-binding proteins or antibodies or paratopes therein bind to human ASGR-1, the antigen-binding proteins or antibodies or paratopes therein are 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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、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、S171、G172, 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 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: 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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: 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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: 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 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: 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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: 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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: 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: 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 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: 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 paratope in the antibody, when bound to human ASGR-1, exhibits antigen-binding activity, e.g., R237, P238, E239, Q240, D242, H257, T259, D260, D261, R263, N265, D267, R271, or Y273 (SEQ ID NO:5). 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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: 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 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: 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 1, 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 binds to human ASGR-1, the antigen binding protein or antibody or paratope in the antibody binds to human ASGR-1 (SEQ ID NO:6). 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 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, 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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, 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein is located within 5 angstroms of at least one of the following residues of 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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: 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: 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 therein is bound to human ASGR-1, the antigen binding protein or antibody or paratope therein 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 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: 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 directed 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 at which the ligand binds to human ASGR-1. In some embodiments, the site 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 site that overlaps with the site 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 present invention provides an isolated antigen binding protein that binds to human ASGR-1 and inhibits human ASGR, ASGR-1 and / or ASGR-2 function, wherein the antigen binding protein does not bind to a variant ASGR-1 protein, wherein the variant ASGR-1 protein is selected from the group consisting of SEQ ID NO: and E280, as set forth in SEQ ID 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 those of skill in the art, such as the method described in Example 7E, infra. In some embodiments, the variant ASGR-1 protein contains 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 set forth 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 K199. 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 at residue W195. In some embodiments, the variant ASGR-1 protein comprises a single mutation at a residue selected from the group consisting of R170, S171, R183, L184, H215, P220, P238, G246, H247, G248, G251, and N265, as set forth 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 at residue R183. In some embodiments, the single mutation is a mutation at residue L184. In some embodiments, the variant ASGR-1 protein comprises a single mutation at a residue selected from the group consisting of P241, D242, D243, Y245, G251, E253, and D260, as set forth 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 selected from the group consisting of or including R170, R237, E239, P241, T259, D260, R263, and N265 as set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 5. In some embodiments, an antigen-binding protein or antibody does not bind to more than one variant ASGR-1 protein, each of which individually comprises a single mutation from 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 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 described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) 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%, or at least about 60% in a patient.

[0021] In some aspects, the present invention provides methods for reducing the risk of developing coronary artery disease or having a myocardial infarction (MI), comprising administering a therapeutically effective dose of an ASGR inhibitor described herein to a patient in need of a reduced 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 described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) 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%, or 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 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 described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) 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 lowering non-HDL cholesterol levels in a patient, comprising administering a therapeutically effective dose of an ASGR inhibitor described herein to a patient in need of lowering 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 described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) 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 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 described herein. In some embodiments, the ASGR, ASGR-1 and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) 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, hi 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 above pre-treatment.

[0025] In some aspects, the present invention provides methods for antagonizing ASGR, ASGR-1, and / or ASGR-2 in a patient, comprising administering a therapeutically effective dose of an ASGR inhibitor 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 described herein. In some embodiments, the ASGR, ASGR-1, and / or ASGR-2 inhibitor is an interfering RNA (e.g., siRNA or shRNA) described herein. [Brief explanation of the drawings]

[0026] [Figure 1A] Figure 1 shows the ASGR-1 sequence alignment of human (SEQ ID NO:32699), cynomolgus monkey (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; the 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). [Figure 2]FIG. 1 shows 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. [Figure 3] Figure 1 provides the human ASGR-1 (SEQ ID NO:32717) vs. human ASGR-2v2 (SEQ ID NO:32718) alignment. [Figure 4]The del12 variant is 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 located 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 the blood of del12 carriers and non-carriers. Arrows indicate both the size of the expected PCR product (239 bp) as well as 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 lacks 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. The sequence around the exon 4-intron 4 boundary (exon 4 sequence in uppercase and intron 4 sequence in lowercase) is 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 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. [Figure 5](A) Del12 variants were typed in the indicated populations, totaling 41,648 CAD cases and 247,374 controls. For each cohort, squares (or diamonds for combined estimates) indicate estimated odds ratios, and lines indicate 95% confidence intervals. There was no evidence of heterogeneity across the eight study populations (Phet = 0.96). (B) Kaplan-Meier curves for survival from a first myocardial infarction in del12 carriers and non-carriers in ASGR-1, stratified by sex. The proportion of individuals without 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] Figure 1.7 Comparison of the relationship 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 reduced expression of human ASGR-1. Panel B is a graphical representation of the relative reduction in human ASGR-1 expression. Panel C demonstrates that CHO cells receiving construct S1662 exhibit 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 reduced 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 in vitro RNAi data in HepG2 cells using construct S1662. Panel A is a Western blot demonstrating the reduction of human ASGR-1 expression. Panel B is a graphical representation of the relative reduction of human ASGR-1 expression. [Figure 11]

[0023] Figure 1 shows in vitro RNAi data in CHO cells transfected with hASGR-2 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]

[0033] Figure 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 of 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 of mouse ASGR-1 expression by various constructs. Panel C is a graphical representation of the relative reduction of mouse ASGR-2 expression by various constructs. [Figure 13]

[0023] Figure 1 shows in vitro RNAi data in HepG2 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 14]

[0023] Figure 1 shows RNAi in vivo data in C57BL / 6J mice using various constructs over a 7-day period, with three injections: one on day 0, one on day 2, and one on day 4. Panel A is a graphical representation of quantitative PCR data showing the relative reduction in mASGR-1 RNA expression in the liver. Panel B is a graphical representation of the relative reduction in mASGR-2 RNA expression in the liver. [Figure 15]

[0023] Figure 1 shows RNAi in vivo data in C57BL / 6J mice using various constructs over a 7-day period, with three injections: one on day 0, one on day 2, and one on day 4. Panel A is a Western blot demonstrating reduced expression of mouse ASGR-1 protein. Panel B is a graphical representation of the relative increase in serum ALP activity. [Figure 16]

[0033] 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 7 days, 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. 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. Panel C shows a close-up of the carbohydrate-binding domain. [Figure 20] 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. 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 irregular carbohydrate-binding loop 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 CBD and 22G5 Fab. Panel B shows a close-up of the ASGR-1 CBD and 22G5 Fab, with dashed lines representing the irregular carbohydrate-binding loop and highlighting the ASGR-1 CBD and indirect inhibition of ligand (GalNAc) binding. Panel B also incorporates a double-headed arrow representing the 5 Å tip-to-tip distance. [Figure 24] FIG. 1 is a representation of the structure of ASGR-1 CBD and 4A2 Fab. [Figure 25] A close-up of the structure of ASGR-1 CBD and 4A2 Fab 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 figure incorporates a double arrow representing the 5 Å tip-to-tip distance. [Figure 26] A close-up of the structure of the ASGR-1 CBD and carbohydrate binding loop with and without 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 dashed lines represent the disordered carbohydrate-binding loop, highlighting the ASGR-1 CBD and its indirect inhibition of ligand (GalNAc) binding. The figure incorporates a double-headed arrow representing a 5 Å tip-to-tip distance. [Figure 29] FIG. 1 is a representation of the structure of ASGR-1 CBD and 4H6 Fab. [Figure 30] A close-up of the structure of the ASGR-1 CBD and 4H6 Fab. The figure depicts the disordered carbohydrate-binding loop with a dashed line, highlighting the ASGR-1 CBD and indirect inhibition of ligand (GalNAc) binding. The figure incorporates a double-headed arrow representing a 5 Å tip-to-tip distance. [Figure 31] FIG. 1 is a representation of the structure of ASGR-1 CBD and 72G9 Fab. [Figure 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 the ligand, highlighting the direct inhibition of ASGR-1 CBD and ligand (GalNAc) binding. [Figure 33] FIG. 1 is a representation of the structure of ASGR-1 CBD and 194A4 Fab. [Figure 34]A close-up of the structure of the ASGR-1 CBD and 194A4 Fab. The figure depicts the disordered carbohydrate-binding loop with a dashed line, highlighting the ASGR-1 CBD and indirect inhibition of ligand (GalNAc) binding. The figure incorporates a double-headed arrow representing a 5 Å tip-to-tip distance. [Figure 35] FIG. 1 is a representation of the structure of ASGR-1 CBD and 54E9 Fab. [Figure 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 54E9 Fan that also overlaps with the structure of the ASGR-1 CBD and the ASGR-1 CBD and ligand, highlighting the 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 show close-ups of the 218G4 Fab structure, which also overlaps with the ASGR-1 CBD and the ASGR-1 CBD and ligand structures. These figures highlight the direct inhibition of ASGR-1 CBD and ligand (GalNAc) binding in the presence of 218G4 Fab. [Figure 39] FIG. 1 is a representation of the structure of ASGR-1 CBD and 176H4 Fab. [Figure 40] A close-up of the structure of 176H4 Fab overlaid with the ASGR-1 CBD and the structures of ASGR-1 and the ligand. This figure highlights the direct inhibition of ASGR-1 CBD and ligand (GalNAc) binding in the presence of 176H4 Fab. [Figure 41] FIG. 1 is a structural depiction of the ASGR-1 CBD and 194C10 Fab, showing the disordered carbohydrate-binding loop with a dashed line, highlighting the ASGR-1 CBD and possible indirect inhibition of ligand (GalNAc) binding. [Figure 42]A close-up of the structure of the ASGR-1 CBD and 194C10 Fab. This figure shows the CDRs of 194C10 interacting with the ASGR-1 CBD, highlighting that these may be direct inhibitors of ASGR-1 CBD and ligand (GalNAc) binding. [Figure 43] Panels AC are graphical representations showing antibody binding results from cells expressing human ASGR-1 and human ASGR-2. [Figure 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. [Figure 45] Panel A is a graphical representation of the effect of the ASGR-1 antibody, 4A2, on serum LDL cholesterol levels in normal cynomolgus monkeys. Panel B is a graphical representation of the effect of the ASGR-1 antibody, 4A2, on serum alkaline phosphatase levels in normal cynomolgus monkeys. Data are expressed as percent change from baseline. [Figure 46] Figure 7E shows a coefficient of determination heat map depicting the coefficient of determination profiles of test ASGR-1 ligand-blocking antibody-reference antibody combinations from arginine / glutamic acid scanning mutagenesis. Dark shading represents highly similar data, while 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 gray). The positions of amino acids identified by Arg / Glu mutation scanning are shown (dark gray 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]

[0023] Figure 2A and Table 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 present 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 a protein alignment of the light and heavy variable regions for certain antigen-binding proteins of the present invention (Table 4). An asterisk "*" indicates a stop codon. Sequences containing a stop codon are represented as different sequences in the sequence listing, although 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 considerations and should be understood to be a single amino acid sequence, unless otherwise stated, as are sequences with one or more stop codons. [Figure 52] 5 is a table presenting a consensus protein alignment of the light and heavy variable regions for certain antigen-binding proteins of the present invention (Table 5). An asterisk "*" indicates a stop codon. Sequences containing a stop codon are represented in the sequence listing as different sequences, although 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 considerations and should be understood to be a single amino acid sequence, unless otherwise stated, as are sequences with one or more stop codons. [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 considerations and should be understood to be single amino acid sequences 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 presenting 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 group 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 specific residues that one skilled in the art may wish to target for genetic engineering. [Figure 57]

[0023] Figure 49 is a group of tables presenting 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] 1 is a graph showing the reliability of protein measurements in cynomolgus monkeys. The Log10 RFU of mean protein levels in the two species is 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 is a graph showing TNFSF8 protein levels in individual animals of different treatment groups across time points. Panel B is a graph showing normalized TNFSF8 protein levels (percentage of time point 0) in individual animals of different treatment groups across time points. Panel C is a graph showing TNFSF8 protein levels in each treatment group (n=3, error bars represent SEM), and panel D is a graph showing the distribution of TNFSF8 protein levels in human ASGR1 del12 carriers and non-carriers. DETAILED DESCRIPTION OF THE INVENTION

[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 a 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 methods 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 present invention further provides compositions, kits, and methods related 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 present invention further provides vectors and plasmids containing such nucleic acids, as well as cells or cell lines containing such nucleic acids and / or vectors and plasmids. Methods provided include, for example, methods for generating, 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 contain 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 of the invention as claimed and are not restrictive. As used herein, the use of the singular includes the plural unless expressly stated otherwise. As used herein, the use of "or" means "and / or" unless expressly stated otherwise. Furthermore, the use of the terms "including" and other forms, such as "includes" and "included," is not limiting. Furthermore, terms such as "element" or "component" encompass both elements and components comprising one unit and elements and components comprising two or more subunits, unless expressly stated otherwise. Furthermore, the use of the term "portion" can include a portion of a moiety or the entire moiety.

[0031] Unless otherwise defined herein, scientific and technical terms used in connection with the present invention shall have the meanings commonly understood by those of ordinary skill in the art. Furthermore, unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular. Generally, 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 the 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); 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 performed according to manufacturer's specifications, as commonly accomplished in the art or as described herein. The terminology used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry described herein are those well known and commonly used in the art. Standard techniques may 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 specified, 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 top 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 position, 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) refers to 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 in which it naturally originates would 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 numerous 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), analogs of DNA or RNA 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 containing a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of the host cell after introduction into the host cell, and thereby are 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., level, timing, or location) of the nucleotide sequence. A "regulatory sequence" is a nucleic acid that affects the expression (e.g., level, timing, or location) 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., polypeptides and / or nucleic acids that bind 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, e.g., a nucleic acid of the invention. A host cell can be a prokaryote, e.g., E. coli, or a eukaryote, e.g., a unicellular eukaryote (e.g., yeast or other fungi), a plant cell (e.g., tobacco or tomato plant cell), an animal cell (e.g., 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 refer to 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 modality that allows for the discovery of associations between disease states and specific targets and / or pathways. For example, genetic analysis of patients with familial hypercholesterolemia led to the discovery of proprotein convertase subtilisin / kexin type 9 (PCSK9), which is involved in regulating serum LDL cholesterol levels and the risk of developing coronary artery disease, ultimately leading to the development of the recently approved anti-hPCSK9 antibody Repatha® (see, e.g., Jackson et al., U.S. Patent No. 8,030,457). Advances in DNA sequencing technology provide a means to sequence the genomes of large numbers of individuals, enabling the discovery of rare variants. deCODE Genetics (Amgen) previously reported a method for analyzing the whole genomes of large numbers of Icelanders to explore associations between genetic variants and traits of interest (Gudbjartsson et al., Nature Genetics; Vol. 47; ( 5 ) Issue; 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 performed 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 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 predicted to generate a truncated ASGR-1 receptor subunit lacking both the oligomerization and extracellular carbohydrate recognition domains (also known as the "CRD," "carbohydrate-binding domain," or "CBD"), or may generate an unstable and rapidly degraded transcript (and therefore no protein) via nonsense-mediated decay. In the present invention, whole-genome sequencing of Icelanders also discovered the second rarest loss-of-function variant in the ASGR-1 gene, a 4-base pair insertion (c.469-472dupAACT or "W158X") in exon 7. This four-base pair insertion in exon 7 causes a frameshift, introducing 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 encode a protein lacking the carbohydrate-recognition domain of the receptor or may generate an unstable, rapidly degraded transcript (thus resulting in no protein) via nonsense-mediated decay. Furthermore, the W158X variant results in all reported refseq transcripts of ASGR-1, regardless of tissue or cell type of expression. While not wishing to be bound by any particular hypothesis, our analysis indicates that del12 and W158X confer lower non-HDL cholesterol levels, protection from CAD and MI, and extended lifespan. Furthermore, our analysis indicates that del12 and W158X are also associated with increased levels of circulating alkaline phosphatase and vitamin B12. Supporting this association between del12 and W158X and increased ALP levels are data from mice carrying the Y272C variant in ASGR-1, which show that such mice exhibit 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 Figure 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, hepatic galactose / N-acetylgalactosamine (GalNAc) receptor, or hepatic lectin receptor. However, this receptor is now more commonly known as "ASGPR" or simply "ASGR."

[0044] ASGR is a C-type lectin expressed on the surface of hepatocytes and is composed of 48 kDa major subunit(s) (ASGR-1) and 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). Receptor complexes can contain homo- and hetero-oligomers of the ASGR-1 and ASGR-2 subunits, with the (ASGR-1)2-(ASGR-2)1 trimer being the most common form and possessing the highest affinity for substrates (Grewal). Other identified forms of ASGR include (ASGR-1)2, (ASGR-1)3, (ASGR-1)2-(ASGR-2)2, and (ASGR-1)3-(ASGR-2)2 (Grewal). Polynucleotide and polypeptide sequences of ASGR-1 and ASGR-2 from several species are known. Table 1 shows the sequences for human, mouse, rat, pig, dog, and cynomolgus monkey. Figures 1A, 1B, and 2 show sequence alignments of ASGR-1 and ASGR-2 from various species, and Figure 3 shows a 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 the removal of sialic acid by sialidase, and thus the term asialoglycoprotein is used to describe the ligand for ASGR. While 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 mentioned above, the W158X variant may generate a transcript that is unstable and rapidly degraded (and therefore does not produce protein) by nonsense-mediated decay.

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

[0047] As used herein, the terms "ASGR, ASGR-1, and / or ASGR-2 function" or "ASGR, ASGR-1, and / or ASGR-2 activity" include 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 with or bind to a ligand. In some embodiments, ASGR function or activity is expressed by the ability of ASGR to interact with or bind 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. 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 the circulation, clearance of IgA from the circulation, removal of apoptotic cells, clearance of low-density lipoprotein (LDL), and / or disposal of cellular fibronectin (Roggenbuck).

[0048] Given 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] ASGR and de-Sinicization RAlthough its ability to mediate the endocytosis and degradation of desialoglyzed glycoproteins has been known for nearly 40 years, the endogenous ligand and physiological function 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 has been reported that ASGR-1- / - mice (lacking any ASGR activity) grow normally and healthily and do not accumulate desialoglyzed glycoproteins in their circulation, but are unable to eliminate 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, which lack obvious phenotypes, the present invention has established a clear physiological role for human ASGR-1 in cardiovascular disease, including but not limited to regulating non-HDL levels and regulating CAD and MI risk.The present invention also demonstrates the association between del12 and W158X and increased levels of circulating ALP and vitamin B12.Furthermore, because heterozygous carriers of del12 live an average of 1.5 years longer than non-carriers, the present invention shows that disrupting one allele of ASGR-1 appears to have an overall beneficial effect.

[0051] Surprisingly, various embodiments provided herein demonstrate that both del12 variant and W158 variant have the opposite effect on non-HDL levels to their effects on ALP and vitamin B12 levels; reducing 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; therefore, ASGR-1 may affect the levels of these molecules through different mechanisms.Since other measures of liver function are not affected, the ALP increase mediated by del12 or W158X is unlikely to reflect underlying liver disease. Both circulating ALP and the vitamin B12 transporter, haptocorrin, are nonsialylated glycoproteins known to bind to 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). While not 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 actively (LDL levels were reduced in these mice) than the asialylated form of 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 have been 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 also directed to the method for inhibiting ASGR function, the method for inhibiting ASGR-1 function and / or the method for inhibiting ASGR-2 function.The present invention is also directed to the molecule (for example, including but not limited to, 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 present invention comprises antigen-binding proteins that bind to ASGR, ASGR-1, and / or ASGR-2 from different species, including, but not limited to, humans, cynomolgus monkeys, pigs, dogs, mice, and rats. In some embodiments, the antigen-binding proteins specifically bind to ASGR, ASGR-1, and / or ASGR-2 from different species, including, but not limited to, humans, cynomolgus monkeys, pigs, dogs, mice, and rats. Exemplary amino acid sequences of human, cynomolgus monkeys, dogs, pigs, rats, 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 comprising 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 comprises at least one CDR from an antibody that binds to the antigen, and in some embodiments, comprises a heavy chain CDR3 from an antibody that binds the antigen. In some embodiments, the antigen-binding fragment comprises all three CDRs from the heavy chain or from the light chain of an antibody that binds the antigen. In further embodiments, the antigen-binding fragment comprises all six CDRs (three from the heavy chain and three from the light chain) from an antibody that binds 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 modifying intact antibodies (e.g., by enzymatic or chemical cleavage) or 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 containing mutations introduced to stabilize the three-dimensional structure of the antigen-binding protein, and fully synthetic scaffolds containing, 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 in conjunction with peptide antibody mimetics ("PAMs").

[0057] Antigen-binding proteins can also include proteins comprising one or more antibody fragments incorporated into a single polypeptide chain or multiple polypeptide chains. For example, antigen-binding proteins 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; see Fredericks et al., Protein Engineering, Design & Selection 17:95-106, 2004 and Powers et al., Journal of Immunological 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 WO 00 / 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; see 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 (C) regions 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)), incorporated herein 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 comprising, from the amino terminus to the 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 comprising, from the amino terminus to the carboxyl terminus, a single immunoglobulin heavy chain variable region (VH), immunoglobulin heavy chain constant domain 1 (CH1), an 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, i.e., IgG1, IgG2, IgG3, and IgG4. The IgA class is further divided into subclasses, i.e., 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). Immunoglobulin heavy chain constant domains can be derived from any immunoglobulin isotype, including subtypes. 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 contains at least two full-length heavy chains and two full-length light chains. The antibody sequence may be derived entirely 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 specified, the term "antibody" also includes antibodies containing two substantially full-length heavy chains and two substantially full-length light chains, provided that 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 one, two, three, four, or five 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 the antibody retains the same or similar binding and / or function as an antibody containing 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, example antigen-binding proteins are described herein in terms of hybridoma lineage number as "number / letter / number" (e.g., 25A4). In such cases, the precise name indicates a specific monoclonal antibody derived from a specific hybridoma having specific light chain variable regions and heavy chain variable regions. In some sections of this disclosure, example 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 the 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] "Polyclonal antibody" refers to a population of antibodies that typically vary widely in composition and binding specificity. "Monoclonal antibody" ("mAb"), as used herein, refers to one or more of a population of antibodies with 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 containing the VL, VH, CL, and CH1 domains; F(ab')2 fragments are bivalent fragments containing two Fab fragments linked by disulfide bridges at the hinge region; Fd fragments contain the VH and CH1 domains; Fv fragments contain the VL and VH domains of a single arm of an antibody; and dAb fragments contain the VH domain, the VL domain, or antigen-binding fragments of the VH or VL domain (U.S. Patent Nos. 6,846,634, 6,696,245, U.S. Patent Application Publication 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. Pat. No. 6,703,199, including fibronectin polypeptide monobodies, and polypeptides disclosed in U.S. Patent Application Publication No. 2005 / 0238646. In some embodiments, the antibody comprises at least one CDR set forth in Table 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 comprising two polypeptide chains, each of which contains a VH and VL domain connected by a linker too short to allow pairing between the two domains on the same chain, so that each domain pairs with a complementary domain on another polypeptide chain (see, e.g., 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 resulting diabody 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, which 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 non-covalently, to make the molecule an antigen-binding protein.

[0068] In some embodiments, antigen-binding proteins of the present invention may incorporate a CDR(s) as part of a larger polypeptide chain, may covalently link a CDR(s) to another polypeptide chain, or may incorporate a 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 a localized surface area. Such a structure may be a naturally occurring polypeptide or polypeptide "fold" (structural motif) or may have one or more modifications, such as amino acid additions, deletions, or substitutions, compared to a naturally occurring polypeptide or fold. Such a scaffold may be derived from a polypeptide of any species (or two or more 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, neocarzinostatin, cytochrome b, CP1 zinc finger, PST1, coiled coil, LACI-D1, Z domain, and tendamistat domain may be used (see, e.g., 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 two or more binding sites are present, the binding sites may be identical to each other or different. For example, an antibody typically has two identical binding sites, while 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, which may be present in 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 the ASGR-1 CBD and inhibits ASGR-1 function in addition to binding to PCSK9 and inhibiting PCSK9 binding to LDLR. Methods for producing bispecific antibodies are known in the art. One such method for producing "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 cells. 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 cells. WO 09 / 089,004, the entire contents of which are incorporated herein by reference.

[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). Human antibodies of the present 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 random or site-specific mutagenesis in vitro or somatic mutation in vivo). Human antibodies can have at least one, two, three, four, five, or more positions replaced with amino acid residues not encoded by human germline immunoglobulin sequences. The definition of human antibody, as used herein, also contemplates fully human antibodies, which contain only non-artificially and / or genetically altered human sequences of antibodies, as can be obtained using techniques or systems known in the art, such as, but not limited to, phage display or transgenic mouse technology.

[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, so 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 an antibody derived from a non-human species. 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, constant domain(s) from a human antibody are fused to 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 the altered amino acid residues are either not critical to the immunospecific binding of the antibody to the antigen, or the changes made to the amino acid sequence are conservative changes, so that binding of the humanized antibody to the antigen is not significantly worse than binding of the non-human antibody to the 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 contain 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, portions of the heavy and / or light chains are 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) are 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 the target molecule to which it binds from performing a biological function. In assessing the binding and / or specificity of an antigen-binding protein, e.g., an antibody or immunologically functional fragment thereof, an antibody or fragment can substantially inhibit binding of a target molecule to its binding partner if excess antibody reduces the amount of 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, inhibition is complete. Measurement of binding reduction can be performed using various assays known to those skilled in the art (e.g., in vitro competitive binding assays), using relevant control molecules to measure actual inhibition. For example, numerous competitive assays are well known in the art, including, but not limited to, the use of competitive ELISA, the BiaCore® platform, the Kinexa® platform, and others. As yet another example, solid phase direct or indirect radioimmunoassays ( RA), solid-phase direct or indirect enzyme immunoassay (EIA), sandwich competitive assay (see, e.g., Stahl 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 assay, solid-phase direct label sandwich assay (see, e.g., Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press); I These include solid-phase direct-labeling RIA using -125 labeling (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, neutralizing ability is characterized and / or described by a competitive assay. In some embodiments, the neutralizing capacity is determined by IC 50 or EC 50The antigen-binding proteins in Table C are described in terms of their potent neutralizing activity. At least the antigen-binding proteins in 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 cells 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 (competing 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 an adjacent epitope that is close enough to the epitope bound by the reference antigen-binding protein (sufficiently close to steric hindrance). Typically, when present in excess, a competing 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 a target protein (e.g., ASGR-1) are mixed with an antigen-binding protein, incubated for a certain period of time, and then washed.These cells are then incubated with a 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 a related control antigen-binding protein indicates inhibition of binding by an antigen-binding protein that blocks or inhibits this binding.

[0078] Another way in which the reduction in binding can be measured is by the half-maximal inhibitory concentration (IC50). IC50 measures the amount or concentration of an antigen-binding protein required to half-inhibit a given property (e.g., ligand binding). 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, antigen binding proteins of the present invention bind to ASGR-1 monomers. In some embodiments, antigen binding proteins of the present invention bind to ASGR-1 oligomers. In further embodiments, antigen binding proteins of the present invention bind to ASGR-2 monomers. In some embodiments, antigen binding proteins of the present invention bind to ASGR-2 oligomers. In certain embodiments, antigen binding proteins of the present invention bind to both ASGR-1 and ASGR-2 monomers. In certain embodiments, antigen binding proteins of the present invention bind to ASGR oligomers comprising (ASGR-1)2-(ASGR-2)1 trimers. In some embodiments, antigen binding proteins of the present invention bind to ASGR oligomers comprising (ASGR-1)2 dimers. In further embodiments, antigen binding proteins of the present invention bind to ASGR oligomers comprising (ASGR-1)3 trimers. In still further embodiments, antigen binding proteins of the present invention bind to ASGR oligomers comprising (ASGR-1)2-(ASGR-2)2 tetramers. In further embodiments, the antigen binding proteins of the invention bind to ASGR oligomers, including (ASGR-1)3-(ASGR-2)2 pentamers. In some embodiments, the antigen binding proteins of the invention bind to multimeric complexes comprising at least two subunits of ASGR-1 and / or ASGR-2.

[0080] In certain embodiments, an antigen-binding protein (e.g., an antibody, 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 desired binding properties of the molecule (e.g., part of the paratope). As used herein, a "paratope" is a location in an antibody that binds to an antigen. The paratope may include several amino acid residues from the VH and / or VL CDRs and may also include residues from the framework region. The paratope binds to an epitope of an antigen. The paratope can 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 a 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, in Example 10, the binding of a representative antigen-binding protein (e.g., an antibody) to human ASGR-1 was analyzed using X-ray crystallography, 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 described 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% or more identity, 95% or more identity, 97% or more identity, or 99% or more identity, or antigen-binding protein variants containing variable domains with 20 or fewer, 15 or fewer, 10 or fewer, or 5 or fewer 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 amino acids within 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 generated with one or more amino acid additions, substitutions, and / or deletions in one or more CDRs that retain 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 a 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. The compound 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 of the 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, and D106 of NO:29170. In some embodiments, the light chain variable region comprises at least one, two, three, four, five, six, seven, 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 one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, 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, SO1, M102, G103, T28, F29, F32, G33, H35, W47, I51, D54, K58, D99, L100, and 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, and 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, and 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. The amino acid sequence includes 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 of the 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 one, two, three, four, five, six, seven, eight, 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 one, two, three, four, five, six, seven, eight, nine, ten, eleven, 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. The compound 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 of the 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, and the light chain variable region is selected from the group consisting of SEQ ID NO:. 27482; and / or the heavy chain variable region comprises at least one, two, three or all 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 1, 2, 3, 4, 5, or all 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 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all 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, 197, Q98, 199, 12, 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, 197, Q98, 199, 12, 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 1, 2, 3, 4, 5, 6, 7, 8, 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 1, 2, 3, 4, 5, 6, 7, 8, 9, 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 among the antigen-binding proteins of the present invention are contemplated. 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 having 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 to the translated IMGT germline V, D, and J genes. First, the V gene was located, followed by the J gene, located in the region downstream of the located V gene, and finally the D gene, located in the region between the V and J regions. Note that because the D gene is a relatively short sequence located in the hypervariable CDR3 region, spurious matching is possible and was taken into account.

[0085] Sequences from each group were then subjected to sequence similarity alignment using a program employing 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 created and then exported as a PDF image, sequence logos were generated in Geneious (v8.1.7, Biomatters). Consensus sequences were generated in Geneious (v8.1.7, Biomatters) using a 0% threshold and exported as a FASTA file. Amino acids that varied within each group are indicated by the symbol X within each consensus sequence. For the consensus sequences resulting from this analysis, see Table 19A VH Consensus 1-14 and Table 20A VL Consensus 1-14 in Figure 55 and Tables 21-48 in Figure 56. In other cases, consensus sequences were generated in Abinitio. See Table 19A VH consensus-15-60 and Table 20A VL consensus 15-54 in Figure 55 and Table 49-134 in Figure 57 for consensus sequences resulting from this analysis.

[0086] Alternatively, different analytical methods readily available to those skilled in the art may be used. For example, consensus sequences can be determined using standard phylogenetic analysis of the CDRs corresponding to the VH (i.e., variable heavy, etc.) and VL (i.e., variable light, etc.) of an antibody. For example, amino acid sequences corresponding to any entire variable domain of a VH or VL can be converted to FASTA format to facilitate comparative alignment and phylogenetic inference. The framework regions of these sequences can then be replaced with artificial linker sequences so that CDRs can be examined alone without introducing any amino acid position weighting bias due to coincident events (e.g., unrelated antibodies that happen to share a common germline framework genetic trait), 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 using a program employing 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. This program also generates phylogenetic trees (phylogenetic tree diagrams) based on sequence similarity alignments using either the UPGMA (unweighted average distance method) or neighbor-joining method (see Saitou and Nei, 1987, Molecular Biology and Evolution 4:406-425) to construct and illustrate the similarities and differences between sequence groups through branch length comparisons and groupings. The resulting original sequence alignment can be used to empirically test and verify 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, regardless of the sequential linkage corresponding to VH or VL within the same sequence. In this approach, a consensus sequence can be determined by aligning each H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 in the group. That is, by aligning the individual H-CDR1 sequences of the antigen-binding proteins, a H-CDR1 consensus sequence can be determined; by aligning the individual H-CDR2 sequences of the antigen-binding proteins, a H-CDR2 consensus sequence can be determined; by aligning the individual H-CDR3 sequences of the antigen-binding proteins, a H-CDR3 consensus sequence can be determined; by aligning the individual L-CDR1 sequences of the antigen-binding proteins, a L-CDR1 consensus sequence can be determined; by aligning the individual L-CDR2 sequences of the antigen-binding proteins, a L-CDR2 consensus sequence can be determined; and by aligning the individual L-CDR3 sequences of the antigen-binding proteins, a L-CDR3 consensus sequence can be determined. Similarities between the sequences within each individual CDR sequence can be identified. Thereby, a consensus sequence of a group of similar sequences within each CDR may be presented.

[0088] Various embodiments of variable heavy chain (VH) consensus amino acid sequences of the invention are set forth in Table 19A of Figure 55 (CDRs are underlined, with CDR1 appearing first). Various embodiments of VH CDR consensus amino acid sequences of the invention are set forth in Tables 19B and 19C of Figure 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) can be present at this position (see Figures 56 and 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 an amino acid is not present at this position (see Figures 56 and 57 for details of the consensus protein alignment). The VH consensus sequences and VH CDR consensus sequences are based on an 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 variable light chain (VL) consensus amino acid sequences of the present invention are set forth in Table 20A of Figure 55 (CDRs are underlined, with CDR1 appearing first). Various embodiments of VL CDR consensus amino acid sequences of the present invention are set forth in Tables 20B and 20C of Figure 55. As noted above, in some cases, an "X" appears in the amino acid sequences set forth in Tables 20A and 20B, indicating that two or more amino acids (or zero amino acids) may be present at this position (see Figures 56 and 57 for details of the consensus protein alignment). In some cases, a "-" appears in Table 20A (which is the result of the consensus alignment) indicating that no amino acid is present at this position (see Figures 56 and 57 for details of the consensus protein alignment). The VL consensus sequence and VL CDR consensus sequence are based on an 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 sequences can, 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 solely to amino acid substitutions, deletions, or additions at any position in the identified consensus sequences (VH, VL, and / or CDRs) with any amino acid contemplated herein.

[0091] In certain embodiments, antigen binding proteins of the invention comprise three VH CDRs and three VL CDRs, wherein 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, wherein 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, wherein 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, wherein 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, wherein 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, wherein 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, wherein 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, wherein 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 within 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 within the VL3 CDR consensus sequence.

[0095] In some embodiments, framework consensus sequences are encompassed by the 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 from 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 from different species.

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

[0098] In some embodiments, the antigen binding proteins of the present invention bind to ASGR-2 from 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 ASGR-2. In some embodiments, the antigen binding proteins of the present invention bind to cynomolgus monkey ASGR-2. In some embodiments, the antigen binding proteins of the present invention bind to pig ASGR-2. In some embodiments, the antigen binding proteins of the present invention bind to dog ASGR-2. In some embodiments, the antigen binding proteins of the present invention bind to mouse ASGR-2. In some embodiments, the antigen binding proteins of the present invention bind to rat ASGR-2. In some embodiments, the antigen binding proteins specifically bind to ASGR-2 from different species.

[0099] In some embodiments, the antigen-binding proteins of the present invention bind to ASGR, ASGR-1, and / or ASGR-2 from two or more different species, and / or bind to ASGR, ASGR-1, and / or ASGR-2 from the same species. For example, but not limited to, antibodies that bind to human and cynomolgus monkey ASGR-1; antibodies that bind to human, cynomolgus monkey, and pig ASGR-1; antibodies that bind to human, cynomolgus monkey, rat, and mouse ASGR-2; antibodies that bind to human ASGR-1 and human ASGR-2; and antibodies that bind to human and cynomolgus monkey ASGR-1 and ASGR-2. In some embodiments, the antigen-binding proteins specifically bind to ASGR, ASGR-1, and / or ASGR-2 from two or more different species, and / or specifically bind 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 also 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 that bind to ASGR, ASGR-1, and / or ASGR-2 and inhibit ligand binding. These properties are contemplated as desirable for reducing or preventing dissociation of the molecule from the receptor during endocytosis to extend the molecule's half-life. In some embodiments, the antigen-binding proteins (e.g., antibodies) exhibit pH-independent binding to their antigens, 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 antigens, such that their affinity for antigen binding under 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 its 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] The asialoglycoprotein receptor ASGR-1 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 structure of the carbohydrate recognition domain ("CRD") 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 previously reported, and when bound to various ligands (e.g., sugars, including but not limited to, lactose, galactose, and / or GalNAc, or glycoproteins displaying 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, antigen binding proteins of the invention bind to the CBD of ASGR-1. In certain embodiments, antigen binding proteins of the invention bind to the CBD of human ASGR-1. In certain embodiments, antigen binding proteins of the invention bind to the CBD of SEQ ID NO:5. In some embodiments, 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 isolated antigen binding proteins that bind to the human ASGR-1 CBD within helix alpha-1 or helix alpha-2. In some embodiments, the invention comprises isolated antigen binding proteins that bind 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 a binding site that is the same as or overlaps with that 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, or other sugars and glycoproteins that can bind to ASGR, ASGR-1, and / or ASGR-2). 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 embodiments, the antigen-binding proteins of the invention bind to the CBD of rat ASGR-1. In some 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 ligand binding to ASGR-1. In specific embodiments, the ligands 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 because 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, antigen-binding proteins of the invention bind to or interact with Y273 of human ASGR-1. In some embodiments, antigen-binding proteins of the invention bind to ASGR-1 at an epitope that includes Y273 of human ASGR-1. In some embodiments, antigen-binding proteins of the invention bind to ASGR-1 at an epitope that renders Y273 of human ASGR-1 unavailable for ligand binding.

[0106] Analysis of the crystal structure of hASGR-1 has revealed specific amino acids involved in the interaction between hASGR-1 and 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). 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, 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, 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 the binding or interaction of a ligand (e.g., a sugar including but not limited to lactose, galactose and / or GalNAc, or a glycoprotein displaying 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, antigen binding proteins of the invention bind to or interact 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 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 some embodiments, antigen binding proteins of the invention bind to or interact with 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, 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 20 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, 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, 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, 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, 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, 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, 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, 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, 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, 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 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 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 20 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, antigen-binding proteins of the invention bind to an epitope that includes at least two 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 an epitope that includes at least three 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 an epitope that includes at least four 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 an epitope that includes at least five 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 an epitope that includes at least six 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 an epitope that includes 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 an epitope that includes 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 an epitope that includes at least nine 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 an epitope that includes at least 10 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 an epitope that includes at least 11 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 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 that displays 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 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., 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 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., a sugar including but not limited to lactose, galactose and / or GalNAc, or a glycoprotein displaying 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 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., a sugar including but not limited to lactose, galactose and / or GalNAc, or a glycoprotein displaying 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 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., 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 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., a sugar including but not limited to lactose, galactose and / or GalNAc, or a glycoprotein displaying such a sugar 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 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., a sugar including but not limited to lactose, galactose and / or GalNAc, or a glycoprotein displaying 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 the 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 the 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 the binding or interaction of a ligand (e.g., lactose, galactose and / or GalNAc) with 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 a further embodiment, the antigen binding proteins of the invention bind to hASGR-1 and block or reduce the 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 the 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 the 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 the 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 the 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 the 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 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 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 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 19 of Q240, D242, W244, E25. 3, 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 a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces the 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 a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces the 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 a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces binding or interaction between a ligand (e.g., lactose, galactose, and / or GalNAc) and at least four of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces binding or interaction between a ligand (e.g., lactose, galactose, and / or GalNAc) and at least five of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces the 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 a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces binding or interaction between a ligand (e.g., lactose, galactose, and / or GalNAc) and at least seven of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces binding or interaction between a ligand (e.g., lactose, galactose, and / or GalNAc) and at least eight of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces binding or interaction between a ligand (e.g., lactose, galactose, and / or GalNAc) and at least nine of Q240, D242, W244, E253, N265, D266, D267, R237, P238, H257, T259, or Y273. In a further embodiment, an antigen binding protein of the invention binds to hASGR-1 and blocks or reduces binding or interaction between a ligand (e.g., lactose, galactose, and / or GalNAc) and 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 the 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 the 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 antigen-binding proteins of the present invention from various characterization bins may be analyzed. For example, antigen-binding proteins of the present invention can be tested 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, 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 containing the same or similar sequence features and patterns will have substantially the same or similar binding characteristics. In specific embodiments, these antigen-binding proteins can bind to the same or similar epitopes with varying affinities.

[0115] Exemplary antigen-binding proteins described herein possess characteristics based on the epitope in ASGR, ASGR-1, and / or ASGR-2 that is 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 that antigen. If the antigen is a protein, the epitope includes specific amino acids that directly contact or interact with the antigen-binding protein. Epitopes can be formed by both contiguous amino acids or noncontiguous amino acids juxtaposed by tertiary folding of the protein. A "linear epitope" is an epitope whose primary amino acid sequence comprises the recognized epitope. A linear epitope typically contains at least three or at least four, more usually at least five, at least six, or at least seven, e.g., about eight to about ten, 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., in a polypeptide, amino acid residues 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). Epitope 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. Generally, an antigen-binding protein specific for a particular target molecule will preferentially recognize an epitope on the target molecule in a complex mixture of proteins and / or macromolecules.

[0117] Methods for characterizing epitopes bound by antigen-binding proteins are well known in the art and include 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, pp. 6935-6942), alanine scanning (Cunningham and Wells, "High-resolution epitope mapping of HGH-receptor interactions by alanine-scanning mutagenesis," Science (1989) 244, pp. 1081-1085), arginine scanning (Lim et al., "A diversity of antibody epitopes can induce signaling through the erythropoietin receptor," Biochemistry (2010) 49, pp. 3797-3804), HD exchange method (Coates et al., "Epitope mapping by amide hydrogen / deuterium exchange coupled with immobilization of antibody, online proteolysis, liquid chromatography and mass spectrometry," Rapid Commun. Mass Spectrom. (2009) 23, pp. 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, "TROSY NMR cross-saturation" Biochemistry (2005) 44, 518-23) and 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 epitopes that are identical or overlapping with the exemplary antigen binding proteins set forth in Tables 2-7. In some embodiments, the antigen binding protein has the same epitope as the exemplary antigen binding protein. In other embodiments, the antigen binding protein binds only the same subset of amino acids as the exemplary antigen binding protein. 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 that is the same as or overlaps with an antigen binding protein in Tables 2-7 and comprises: a) a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of an antigen binding protein set forth in Tables 2-7; b) a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of an antigen binding protein set forth in Tables 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 present invention have an epitope that is identical to or overlaps 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, or identical to the amino acid sequence of 4H6; a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 4H6; and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 4H6; a light chain variable domain that is at least 90% identical, at least 95% identical, or 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 that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 22G5, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 22G5; a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 194A4, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 194A4; a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 218G4G4, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 18G4; a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 176H4, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 176H4; a light chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 194C10, and a heavy chain variable domain that is at least 90% identical, at least 95% identical, or identical to the amino acid sequence of 194C10;

[0121] In certain embodiments, the ASGR-1 antigen binding proteins of the present invention have an epitope that is identical to or overlaps with an antibody in Tables 2-7 and comprise a light chain variable domain comprising: an LCDR1 having no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in Table 2; an LCDR2 having no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in Table 2; and an LCDR3 having no more than three 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 three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in Table 2; an HCDR2 having no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in Table 2; and an HCDR3 having no more than three 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 that is identical to or overlaps with an antibody in Table A, B, C, or 6, and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from an LCDR1 sequence set forth in Table A, B, C, or 6; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from an LCDR2 sequence set forth in Table A, B, C, or 6; and an LCDR3 with no more than three amino acid additions, deletions, or substitutions from an LCDR3 sequence set forth in Table A, B, C, or 6; and a heavy chain variable domain comprising: a) an HCDR1 with no more than three amino acid additions, deletions, or substitutions from an HCDR1 sequence set forth in Table A, B, C, or 6; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from an HCDR2 sequence set forth in Table A, B, C, or 6; and an HCDR3 with no more than three amino acid additions, deletions, or substitutions from an 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 that is identical to or overlaps with that of antibody 25A4 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:480; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8492; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4488; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12500; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 4H6 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:894; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8906; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4902; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12914; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 4A2 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:1130; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:9142; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:5136; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:13148; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 5E5 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:974; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8986; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4982; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12994; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 7E11 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:872; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8884; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4880; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12892; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 54E9 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:3448; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:11460; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:7452; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:15464; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 22G5 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:326; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:8338; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:4334; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:12346; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 194A4 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:2780; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:10792; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:6786; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:14798; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 218G4 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:3746; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:11758; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:7750; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:15762; and an HCDR3 with no more than three 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 that is the same as or overlaps with that of antibody 176H4 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:2502; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:10514; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:6508; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:14520; and an HCDR3 with no more than three 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 that is identical to or overlaps with that of antibody 194C10 and comprises a light chain variable domain comprising: an LCDR1 with no more than three amino acid additions, deletions, or substitutions from the LCDR1 sequence set forth in SEQ ID NO:2792; an LCDR2 with no more than three amino acid additions, deletions, or substitutions from the LCDR2 sequence set forth in SEQ ID NO:10804; and an LCDR3 with no more than three 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 with no more than three amino acid additions, deletions, or substitutions from the HCDR1 sequence set forth in SEQ ID NO:6798; an HCDR2 with no more than three amino acid additions, deletions, or substitutions from the HCDR2 sequence set forth in SEQ ID NO:14810; and an HCDR3 with no more than three 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, antigen-binding proteins of the invention (e.g., antibodies or antibody fragments thereof) compete with antigen-binding proteins listed in Tables 2-7. In some embodiments, antigen-binding proteins of the invention (e.g., antibodies or antibody fragments thereof) compete with antigen-binding proteins listed in Tables A, B, and C. In some embodiments, antigen-binding proteins of the invention (e.g., antibodies or antibody fragments thereof) compete with antigen-binding proteins listed in Table 6. "Compete" or "compete" means that antigen-binding proteins compete for the same epitope or binding site on a target. Such competition can be determined by assays in which a reference antigen-binding protein (e.g., an antibody or antibody fragment) prevents or inhibits specific binding of a 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., Stahl 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 Patients"). (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 can be similarly affected by mutagenesis of ASGR, ASGR-1, and / or ASGR-2. Briefly, 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 multiple individual mutations, residues that play a direct role in binding or are sufficiently close to the antibody that 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 comprise the binding epitope 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). Generally, arginine and glutamic acid are substituted (typically individually) for amino acids in the wild-type polypeptide because these amino acids are charged and bulky, potentially disrupting 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 these 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 SEQ ID NO:6. As shown in NO:5, they are 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 mutations at 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 mutations at 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 mutations at 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 mutation of 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 mutation of R183. In some embodiments, the ASGR-1 antigen binding protein shares the properties of antibody 17H6, and its binding is inhibited by any mutation of 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 mutation of 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 properties 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 properties 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 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 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 group 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 predetermined distance from 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, the 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, 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, 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, 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 includes 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) is 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 between 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 between antibody 72G9 and a ligand (e.g., GalNAc), including those in which any of Q240, D242, or W244 (SEQ ID NO:5) is present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface between antibody 72G9 and a ligand (e.g., GalNAc), including those in which any of Q240, D242, W244, E239, P241, D243, Y245, G246, or G252 (SEQ ID NO:5) is present within the interface. In some embodiments, the ASGR-1 antigen-binding protein forms an interface with ASGR-1, which overlaps with the interface between the antibody 72G9 and the ligand (e.g., GalNAc), including those in which any of Q240, D242, W244, R237, or E253 (SEQ ID NO: 5) is present within the interface. As described in the Examples below, the degree of inhibition caused by 72G9 is lower than that of 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 the ASGR-1 protein and the antibody when bound to each other. For example, when the 72G9 antibody is bound to ASGR-1, there is still enough space (albeit less) for the ligand to reach the binding site.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 between antibody 54E9 and a ligand (e.g., GalNAc), including one in which any of N209, R237, Q240, D242, H257, T259, N265, D267, or Y273 (SEQ ID NO:5) is present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface between antibody 54E9 and a ligand (e.g., GalNAc), including one in which any of N209, R237, Q240, D242, H257, T259, N265, D267, Y273, P238, E239, D260, R263, or R271 (SEQ ID NO:5) is present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface between 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) is present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface between 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) is present within the interface.In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface between antibody 218G4 and a ligand (e.g., GalNAc), including one in which any of N209, H257, N265, D267, or Y273 (SEQ ID NO:5) is present within the interface. In some embodiments, the ASGR-1 antigen binding protein forms an interface with ASGR-1 that overlaps with the interface between 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) is 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, ASGR-1 antigen binding proteins form an interface with ASGR-1 that overlaps with the interface between 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) is present within the interface. In some embodiments, ASGR-1 antigen binding proteins form an interface with ASGR-1 that overlaps with the interface between 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) is 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 crystallography, as described in Example 10. Binding interactions between huASGR-1 and various embodiments of antigen binding proteins (e.g., antibodies) of the invention were also assessed using methodologies including epitope binning, as described in Example 7D, and arginine / glutamic acid mutation profiling, as described in Example 7E. A summary of the data obtained by these methodologies is provided in Table D below. This summary illustrates the various binding characteristics of representative antigen binding proteins (e.g., antibodies)...

Claims

1. 1. A pharmaceutical composition for reducing the risk of cardiovascular disease in a patient in need thereof comprising a therapeutically effective dose of an inhibitor of ASGR-1 (asialoglycoprotein receptor 1), wherein the inhibitor is an interfering RNA that reduces expression of ASGR-1, or an isolated antigen binding protein that binds to human ASGR-1 and inhibits binding between human ASGR-1 and GalNAc residues of asialylated glycoproteins, thereby reducing LDL cholesterol levels.

2. The pharmaceutical composition according to claim 1 , wherein the cardiovascular disease is coronary artery disease or myocardial infarction.

3. A pharmaceutical composition for lowering LDL cholesterol levels in a patient in need thereof, comprising a therapeutically effective dose of an inhibitor of ASGR-1, wherein the inhibitor is an interfering RNA that reduces expression of ASGR-1, or an isolated antigen binding protein that binds to human ASGR-1 and inhibits binding between human ASGR-1 and GalNAc residues of asialylated glycoproteins, thereby lowering LDL cholesterol levels.

4. A pharmaceutical composition for lowering non-HDL cholesterol levels in a patient in need thereof comprising a therapeutically effective dose of an inhibitor of ASGR-1, wherein the inhibitor is an interfering RNA that reduces expression of ASGR-1, or an isolated antigen binding protein that binds to human ASGR-1 and inhibits binding between human ASGR-1 and GalNAc residues of non-sialylated glycoproteins.

5. A pharmaceutical composition for increasing serum ALP (alkaline phosphatase) levels in a patient, comprising a therapeutically effective dose of an inhibitor of ASGR-1, wherein the inhibitor is an interfering RNA that reduces expression of ASGR-1, or an isolated antigen-binding protein that binds to human ASGR-1 and inhibits binding between human ASGR-1 and a GalNAc residue of serum ALP.

6. The pharmaceutical composition of any one of claims 1 to 5, wherein the inhibitor is an interfering RNA that reduces the expression of ASGR-1.

7. The pharmaceutical composition of any one of claims 1 to 5, wherein the inhibitor is the isolated antigen binding protein.

8. 8. The pharmaceutical composition of claim 7, for simultaneous or sequential administration with at least one agent that reduces cholesterol.

9. 9. The pharmaceutical composition of claim 8, wherein the at least one pharmaceutical agent is a statin, an anti-PCSK9 inhibitor, or a combination thereof.

10. 10. The pharmaceutical composition of claim 9, wherein the at least one agent 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.

11. A pharmaceutical composition comprising an interfering RNA of human ASGR-1 (asialoglycoprotein receptor 1) for reducing the risk of cardiovascular disease in a patient in need thereof.

12. 12. The pharmaceutical composition of claim 11, which is at least one of the following: (a) the cardiovascular disease is coronary artery disease or myocardial infarction; (b) the pharmaceutical composition is for treating the cardiovascular disease; or (c) The pharmaceutical composition is for preventing the cardiovascular disease.

13. A pharmaceutical composition comprising an interfering RNA of human ASGR-1 (asialoglycoprotein receptor 1) for lowering LDL (low density lipoprotein) cholesterol levels in a patient in need thereof.

14. (a) the patient has or is at risk of having coronary artery disease or is at risk of having a myocardial infarction, and / or (b) the pharmaceutical composition is for reducing LDL cholesterol levels by at least 20%; The pharmaceutical composition of claim 13.

15. A pharmaceutical composition comprising an interfering RNA of human ASGR-1 (asialoglycoprotein receptor 1) for lowering non-HDL (non-high density lipoprotein) cholesterol levels in a patient in need thereof.

16. (a) the patient has or is at risk of having coronary artery disease or is at risk of having a myocardial infarction, and / or (b) the pharmaceutical composition is for reducing non-HDL cholesterol levels by at least 20%; 16. The pharmaceutical composition of claim 15.

17. A pharmaceutical composition comprising an interfering RNA of human ASGR-1 (asialoglycoprotein receptor 1) for increasing serum ALP (alkaline phosphatase) levels in a patient.

18. The pharmaceutical composition of claim 17, wherein the pharmaceutical composition is for increasing serum ALP levels by at least 40%.

19. A pharmaceutical composition according to any one of claims 11 to 18, which is at least one of the following: (a) the interfering RNA comprises or is an siRNA or shRNA; (b) the interfering RNA is at least partially double-stranded; or (c) The pharmaceutical composition is for reducing the expression of ASGR-1 (asialoglycoprotein receptor 1).

20. A pharmaceutical composition according to any one of claims 11 to 19, which is at least one of the following: (a) the interfering RNA is an siRNA comprising a sense strand and an antisense strand, the antisense strand comprising a region having a sequence complementary to a human ASGR-1 (asialoglycoprotein receptor 1) sequence, the region comprising at least 15 contiguous nucleotides; (b) the interfering RNA comprises a sense strand and an antisense strand, the sense strand comprising a sequence substantially complementary to a sequence of the antisense strand, and forming a duplex region of 15-30 base pairs in length; (c) the interfering RNA comprises a sense strand and an antisense strand, the sense strand comprises a sequence complementary to the sequence of the antisense strand, and forms a duplex region of 15 to 30 base pairs in length; (d) the interfering RNA comprises a duplex region of 19 to 25 base pairs in length; (e) the interfering RNA is 15-30 nucleotides in length and comprises a sequence 100% complementary to a sequence in human ASGR-1 (asialoglycoprotein receptor 1), the sequence in human ASGR-1 (asialoglycoprotein receptor 1) being 15-30 nucleotides in length; (f) the interfering RNA is an siRNA comprising a sense strand of 15-30 nucleotides and an antisense strand of 15-30 nucleotides; or (g) the interfering RNA is an siRNA comprising a sense strand of 19 to 25 nucleotides and an antisense strand of 19 to 25 nucleotides;

21. A pharmaceutical composition according to any one of claims 11 to 20, which is at least one of the following: (a) the interfering RNA is conjugated to a targeting moiety, the targeting moiety targeting cells of the liver; (b) the interfering RNA comprises modified nucleotides, or (c) the pharmaceutical composition is for parenteral administration.

22. The pharmaceutical composition according to any one of claims 11 to 12, which is for administration simultaneously or sequentially with at least one drug that reduces cholesterol.

23. 23. The pharmaceutical composition of claim 22, wherein the at least one pharmaceutical agent is a statin, an anti-PCSK9 inhibitor, or a combination thereof.

24. 1. A pharmaceutical composition comprising an siRNA targeting human ASGR-1 (asialoglycoprotein receptor 1) for reducing the risk of cardiovascular disease or myocardial infarction in a patient in need thereof, wherein the siRNA comprises a sense strand that is 15-30 nucleotides in length and an antisense strand that is 15-30 nucleotides in length, the antisense strand comprises a sequence that is complementary to a human ASGR1 sequence, and the sense strand comprises a sequence that is complementary to a sequence of the antisense strand, thereby forming a duplex that is 15-30 base pairs in length.

25. 1. A pharmaceutical composition comprising an siRNA targeting human ASGR-1 (asialoglycoprotein receptor 1) for increasing serum ALP (alkaline phosphatase) levels in a patient, wherein the siRNA comprises a sense strand that is 15-30 nucleotides in length and an antisense strand that is 15-30 nucleotides in length, the antisense strand comprises a sequence that is complementary to a human ASGR1 sequence, and the sense strand comprises a sequence that is complementary to a sequence of the antisense strand, thereby forming a duplex that is 15-30 base pairs in length.

26. 21. The pharmaceutical composition of claim 20, wherein the interfering RNA is as described in (e) and further wherein the ASGR-1 (asialoglycoprotein receptor 1) is human ASGR-1 having the sequence of SEQ ID NO: 32699.

27. 27. The pharmaceutical composition of claim 26, wherein the interfering RNA consists essentially of a sense strand that is 15-30 nucleotides in length and an antisense strand that is 15-30 nucleotides in length.

28. 28. The pharmaceutical composition of claim 27, wherein the sense strand is at least 90% complementary to the antisense strand, and the duplex formed by the sense strand and the antisense strand is 15-30 base pairs in length.

29. The pharmaceutical composition of claims 11, 13, 15, and 17, wherein the ASGR-1 is human ASGR-1 having the sequence of SEQ ID NO: 32699.

30. 30. The pharmaceutical composition of claim 29, wherein the interfering RNA is an siRNA or an shRNA.

31. 31. The pharmaceutical composition of claim 29 or 30, wherein the interfering RNA comprises modified nucleotides.

32. The pharmaceutical composition of any one of claims 29 to 31, wherein the interfering RNA is conjugated to a targeting moiety.

33. 25. The pharmaceutical composition of claim 24, wherein the cardiovascular disease is coronary artery disease.

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