Human Anti-OPGL Neutralizing Antibodies As Selective OPGL Pathway Inhibitors

Abstract

Monoclonal antibodies and hybridomas producing them that interact with osteoprotegerin ligand (OPGL) are provided. Methods of treating osteopenic disorders by administering a pharmaceutically effective amount of antibodies to OPGL are also provided. Methods of detecting the amount of OPGL in a sample using antibodies to OPGL are further provided.

1 . An isolated human antibody that specifically binds osteoprotegerin ligand (OPGL), comprising a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region comprising an amino acid sequence as set forth in any of SEQ ID NO: 6, SEQ ID NO: 14, SEQ ID NO: 22, or SEQ ID NO: 26, or an antigen-binding or an immunologically functional immunoglobulin fragment thereof. 
     
     
         2 .- 20 . (canceled) 
     
     
         21 . An isolated human antibody that specifically binds osteoprotegerin ligand (OPGL), wherein the antibody comprises:
 a. a heavy chain having a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 6, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain having a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 8, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof   b. a heavy chain having a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 14, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain having a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 16, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof   c. a heavy chain having a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 22, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain having a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 24, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof;   d. a heavy chain having a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 26, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain having a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 28, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof;   e. a heavy chain comprising an amino acid sequence as set forth in SEQ ID NO: 30, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain comprising an amino acid sequence as set forth in SEQ ID NO: 32, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof;   f. a heavy chain comprising an amino acid sequence as set forth in SEQ ID NO: 38, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain comprising an amino acid sequence as set forth in SEQ ID NO: 40, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof;   g. a heavy chain comprising an amino acid sequence as set forth in SEQ ID NO: 46, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain comprising an amino acid sequence as set forth in SEQ ID NO: 48, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof; or   h. a heavy chain comprising an amino acid sequence as set forth in SEQ ID NO: 50, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof, and a light chain comprising an amino acid sequence as set forth in SEQ ID NO: 52, an antigen-binding fragment thereof, or an immunologically functional immunoglobulin fragment thereof.   
     
     
         22 .- 49 . (canceled) 
     
     
         50 . The antibody of any of  claims 1  or  21 , wherein the antibody inhibits binding of OPGL to an osteoclast differentiation and activation receptor (ODAR). 
     
     
         51 . A method of treating an osteopenic disorder in a patient, comprising administering to a patient a pharmaceutically effective amount of the antibody of  claim 50 . 
     
     
         52 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of the antibody of  claim 50 . 
     
     
         53 . A method of treating an osteopenic disorder in a patient; comprising administering to a patient the pharmaceutical composition of  claim 52 . 
     
     
         54 . A method for detecting OPGL in a biological sample comprising:
 (a) contacting the sample with the antibody of any of  claims 1  or  21 ; under conditions that allow for binding of the antibody to OPGL;   (b) and measuring the level of bound antibody in the sample.   
     
     
         55 .- 73 . (canceled) 
     
     
         74 . An isolated nucleic acid molecule that encodes an antibody or antigen binding fragment thereof, that specifically binds osteoprotegerin ligand (OPGL), wherein the antibody or antigen binding fragment comprises: SEQ ID NO: 6; SEQ ID NO: 8; SEQ ID NO: 10; SEQ ID NO: 12; SEQ ID NO: 14; SEQ ID NO: 16; SEQ ID NO: 18; SEQ ID NO: 20; SEQ ID NO: 22; SEQ ID NO: 24; SEQ ID NO: 26; or SEQ ID NO: 28. 
     
     
         75 . An isolated nucleic acid molecule that encodes an antibody or antigen binding fragment thereof, that specifically binds osteoprotegerin ligand (OPGL), wherein the antibody or antigen binding fragment comprises: SEQ ID NO: 30; SEQ ID NO: 32; SEQ ID NO: 34; SEQ ID NO: 36; SEQ ID NO: 38; SEQ ID NO: 40; SEQ ID NO: 42; SEQ ID NO: 44; SEQ ID NO: 46; SEQ ID NO: 48; SEQ ID NO: 50; or SEQ ID NO: 52. 
     
     
         76 . An isolated nucleic acid molecule that encodes an antibody or antigen binding fragment thereof, that specifically binds osteoprotegerin ligand (OPGL), wherein the antibody or antigen binding fragment thereof comprises:
 (a) SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 92, SEQ ID NO: 93, and SEQ ID NO: 94;   (b) SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 95, SEQ ID NO: 93, and SEQ ID NO: 94;   (c) SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 82, SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 98;   (d) SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 96, SEQ ID NO: 97, and SEQ ID NO: 98;   (e) SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 99, SEQ ID NO: 97, and SEQ ID NO: 100; or   (f) SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 101, SEQ ID NO: 97, and SEQ ID NO: 102.   
     
     
         77 . An isolated nucleic acid molecule that comprises a sequence that is complementary to the nucleic acid molecule of any of  claims 74 - 76 . 
     
     
         78 . An expression vector comprising a nucleic acid molecule of any of  claims 74 - 76 . 
     
     
         79 . An expression vector comprising a nucleotide sequence comprising two nucleic molecules of any of  claims 74 - 75 . 
     
     
         80 . The expression vector of  claim 79 , wherein the two nucleic acid molecules encode amino acid sequences comprising:
 (a) SEQ ID NO: 6 and SEQ ID NO: 8;   (b) SEQ ID NO: 10 and SEQ ID NO: 12;   (c) SEQ ID NO: 14 and SEQ ID NO: 16;   (d) SEQ ID NO: 18 and SEQ ID NO: 20;   (e) SEQ ID NO: 22 and SEQ ID NO: 24;   (f) SEQ ID NO: 25 and SEQ ID NO: 28;   (g) SEQ ID NO: 30 and SEQ ID NO: 32;   (h) SEQ ID NO: 34 and SEQ ID NO: 36;   (i) SEQ ID NO: 38 and SEQ ID NO: 40;   (j) SEQ ID NO: 42 and SEQ ID NO: 44;   (k) SEQ ID NO: 46 and SEQ ID NO: 48; or   (l) SEQ ID NO: 50 and SEQ ID NO: 52.   
     
     
         81 . An isolated host cell comprising the expression vector of  claim 78 . 
     
     
         82 . An isolated host cell comprising the expression vector of  claim 79 . 
     
     
         83 . An isolated host cell comprising a first expression vector and a second expression vector each according to  claim 78 , wherein the first and second expression vectors each comprise a different sequence encoding amino acid sequences selected from the combinations:
 (a) SEQ ID NO: 6 and SEQ ID NO: 8;   (b) SEQ ID NO: 10 and SEQ ID NO: 12;   (c) SEQ ID NO: 14 and SEQ ID NO: 16;   (d) SEQ ID NO: 18 and SEQ ID NO: 20;   (e) SEQ ID NO: 22 and SEQ ID NO: 24;   (f) SEQ ID NO: 25 and SEQ ID NO: 28;   (g) SEQ ID NO: 30 and SEQ ID NO: 32;   (h) SEQ ID NO: 34 and SEQ ID NO: 36;   (i) SEQ ID NO: 38 and SEQ ID NO: 40;   (j) SEQ ID NO: 42 and SEQ ID NO: 44;   (k) SEQ ID NO: 46 and SEQ ID NO: 48; or   (l) SEQ ID NO: 50 and SEQ ID NO: 52.   
     
     
         84 . The host cell of  claim 81  wherein the host cell is a eukaryotic cell. 
     
     
         85 . The host cell of  claim 83  wherein the host cell is a eukaryotic cell. 
     
     
         86 . The host cell of  claim 85 , wherein the eukaryotic cell is a CHO cell. 
     
     
         87 . A cell line comprising the expression vector of  claim 78 . 
     
     
         88 . A process of producing an antibody or antigen binding fragment thereof; comprising culturing the host cell of  claim 81  under suitable conditions to express the antibody or antigen binding fragment thereof; and optionally isolating the antibody or antigen binding fragment thereof from the culture, wherein the antibody or antigen binding fragment thereof specifically binds human osteoprotegerin ligand (OPGL). 
     
     
         89 . An antibody or antigen binding fragment thereof produced by the process of  claim 87 .