Anti-Glypican 3 Antibody Having Modified Sugar Chain

Abstract

An anti-glypican 3 antibody with modified sugar chains, more specifically, an anti-glypican 3 antibody lacking fucose is provided. The anti-glypican 3 antibody with modified sugar chains of the present invention may be produced by a process comprising introducing a nucleic acid encoding an anti-glypican 3 antibody into host cells with reduced fucose addition capability, such as YB2/0 cells and cells lacking a fucose transporter. The anti-glypican 3 antibody with modified sugar chains of the present invention has a high level of cytotoxic activity and therefore is useful as a cell growth inhibitor such as an anticancer agent.

1 . A composition comprising an anti-glypican 3 antibody, wherein the anti-glypican 3 antibody comprises a heavy chain variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:25, a CDR2 comprising the amino acid sequence of SEQ ID NO:26, and a CDR3 comprising the amino acid sequence of SEQ ID NO:27, wherein the sugar chain component of the antibody has been modified. 
     
     
         2 . A composition comprising an anti-glypican 3 antibody, wherein the anti-glypican 3 antibody comprises a light chain variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:45, a CDR2 comprising the amino acid sequence of SEQ ID NO:46, and a CDR3 comprising the amino acid sequence of SEQ ID NO:60, wherein the sugar chain component of the antibody has been modified. 
     
     
         3 . The composition of  claim 1 , wherein the anti-glypican 3 antibody further comprises a light chain variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:45, a CDR2 comprising the amino acid sequence of SEQ ID NO:46, and a CDR3 comprising the amino acid sequence of SEQ ID NO:60. 
     
     
         4 . A composition comprising an anti-glypican 3 antibody, wherein the anti-glypican 3 antibody comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, and SEQ ID NO:87, and wherein the sugar chain component of the antibody has been modified. 
     
     
         5 . A composition comprising an anti-glypican 3 antibody, wherein the anti-glypican 3 antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:109, and wherein the sugar chain component of the antibody has been modified. 
     
     
         6 . The composition of  claim 4 , wherein the anti-glypican 3 antibody further comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:109. 
     
     
         7 . A composition comprising an anti-glypican 3 antibody, wherein the anti-glypican 3 antibody comprises a light chain variable region comprising a CDR2 comprising the amino acid sequence of SEQ ID NO:46, a CDR3 comprising the amino acid sequence of SEQ ID NO:60, and a CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, and SEQ ID NO:104. 
     
     
         8 . The composition of  claim 7 , wherein the anti-glypican 3 antibody comprises an amino acid sequence selected from the group consisting of SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, and SEQ ID NO:121. 
     
     
         9 . The composition of  claim 1 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         10 . The composition of  claim 2 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         11 . The composition of  claim 3 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         12 . The composition of  claim 4 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         13 . The composition of  claim 5 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         14 . The composition of  claim 6 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         15 . The composition of  claim 7 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         16 . The composition of  claim 8 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 20% of all anti-glypican 3 antibodies in the composition. 
     
     
         17 . The composition of  claim 1 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         18 . The composition of  claim 2 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         19 . The composition of  claim 3 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         20 . The composition of  claim 4 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         21 . The composition of  claim 5 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         22 . The composition of  claim 6 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         23 . The composition of  claim 7 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         24 . The composition of  claim 8 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 50% of all anti-glypican 3 antibodies in the composition. 
     
     
         25 . The composition of  claim 1 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         26 . The composition of  claim 2 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         27 . The composition of  claim 3 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         28 . The composition of  claim 4 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         29 . The composition of  claim 5 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         30 . The composition of  claim 6 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         31 . The composition of  claim 7 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         32 . The composition of  claim 8 , wherein the sugar chain component has been modified to be fucose-deficient and wherein fucose-deficient anti-glypican 3 antibodies comprise more than 90% of all anti-glypican 3 antibodies in the composition. 
     
     
         33 . A method for producing an anti-glypican 3 antibody with a modified sugar chain component, the method comprising:
 (a) providing a cell expressing a nucleic acid encoding an anti-glypican 3 antibody, wherein the cell is a mutant cell that has a reduced capability of adding fucose to sugar chains as compared to a corresponding wild type cell; and   (b) culturing the mutant cell, thereby producing the anti-glypican 3 antibody with a modified sugar chain component, wherein the anti-glypican 3 antibody comprises a heavy chain variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:25, a CDR2 comprising the amino acid sequence of SEQ ID NO:26, and a CDR3 comprising the amino acid sequence of SEQ ID NO:27.   
     
     
         34 . A method for producing an anti-glypican 3 antibody with a modified sugar chain component, the method comprising:
 (a) providing a cell expressing a nucleic acid encoding an anti-glypican 3 antibody, wherein the cell is a mutant cell that has a reduced capability of adding fucose to sugar chains as compared to a corresponding wild type cell; and   (b) culturing the mutant cell, thereby producing the anti-glypican 3 antibody with a modified sugar chain component, wherein the anti-glypican 3 antibody comprises a light chain variable region comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:45, a CDR2 comprising the amino acid sequence of SEQ ID NO:46, and a CDR3 comprising the amino acid sequence of SEQ ID NO:60.   
     
     
         35 . A method for producing an anti-glypican 3 antibody with a modified sugar chain component, the method comprising:
 (a) providing a cell expressing a nucleic acid encoding an anti-glypican 3 antibody, wherein the cell is a mutant cell that has a reduced capability of adding fucose to sugar chains as compared to a corresponding wild type cell; and   (b) culturing the mutant cell, thereby producing the anti-glypican 3 antibody with a modified sugar chain component, wherein the anti-glypican 3 antibody comprises a light chain variable region comprising a CDR2 comprising the amino acid sequence of SEQ ID NO:46, a CDR3 comprising the amino acid sequence of SEQ ID NO:60, and a CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, and SEQ ID NO:104.   
     
     
         36 . The method of  claim 33 , wherein the mutant cell lacks a fucose transporter. 
     
     
         37 . The method of  claim 34 , wherein the mutant cell lacks a fucose transporter. 
     
     
         38 . The method of  claim 35 , wherein the mutant cell lacks a fucose transporter. 
     
     
         39 . The method of  claim 33 , further comprising introducing the nucleic acid encoding the anti-glypican 3 antibody into the mutant cell. 
     
     
         40 . The method of  claim 34 , further comprising introducing the nucleic acid encoding the anti-glypican 3 antibody into the mutant cell. 
     
     
         41 . The method of  claim 35 , further comprising introducing the nucleic acid encoding the anti-glypican 3 antibody into the mutant cell. 
     
     
         42 . The method of  claim 33 , further comprising collecting and purifying the antibody. 
     
     
         43 . The method of  claim 34 , further comprising collecting and purifying the antibody. 
     
     
         44 . The method of  claim 35 , further comprising collecting and purifying the antibody. 
     
     
         45 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 1  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         46 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 2  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         47 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 3  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         48 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 4  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         49 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 5  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         50 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 6  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         51 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 7  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         52 . A method of inhibiting cell growth, the method comprising contacting the composition of  claim 8  with mammalian cells in an amount sufficient to inhibit growth of the cells. 
     
     
         53 . A method of treating cancer, the method comprising administering the composition of  claim 1  to a patient in need of treatment for cancer. 
     
     
         54 . A method of treating cancer, the method comprising administering the composition of  claim 2  to a patient in need of treatment for cancer. 
     
     
         55 . A method of treating cancer, the method comprising administering the composition of  claim 3  to a patient in need of treatment for cancer. 
     
     
         56 . A method of treating cancer, the method comprising administering the composition of  claim 4  to a patient in need of treatment for cancer. 
     
     
         57 . A method of treating cancer, the method comprising administering the composition of  claim 5  to a patient in need of treatment for cancer. 
     
     
         58 . A method of treating cancer, the method comprising administering the composition of  claim 6  to a patient in need of treatment for cancer. 
     
     
         59 . A method of treating cancer, the method comprising administering the composition of  claim 7  to a patient in need of treatment for cancer. 
     
     
         60 . A method of treating cancer, the method comprising administering the composition of  claim 8  to a patient in need of treatment for cancer. 
     
     
         61 . The method of  claim 53 , wherein the cancer is liver cancer. 
     
     
         62 . The method of  claim 54 , wherein the cancer is liver cancer. 
     
     
         63 . The method of  claim 55 , wherein the cancer is liver cancer. 
     
     
         64 . The method of  claim 56 , wherein the cancer is liver cancer. 
     
     
         65 . The method of  claim 57 , wherein the cancer is liver cancer. 
     
     
         66 . The method of  claim 58 , wherein the cancer is liver cancer. 
     
     
         67 . The method of  claim 59 , wherein the cancer is liver cancer. 
     
     
         68 . The method of  claim 60 , wherein the cancer is liver cancer.