Human antibodies that have mn binding and cell adhesion-neutralizing activity

Abstract

The invention is composed of monoclonal human MN antibodies or MN antibody fragments that target the GEEDLP repeat within the proteoglycan domain. The proteoglycan domain of the MN cell surface protein contains four of these identical GEEDLP repeats. Binding to the desired epitope is verified by competition ELISA, where ELISA signal can be attenuated by co-incubation with a peptide containing this repeat (PGEEDLPGEEDLP). This inhibition of binding can also be verified using Biacore assays, where binding of desired antibodies to immobilized MN or proteoglycan peptides can be inhibited by the peptide repeat. In addition to binding to the peptide repeat, human anti-MN antibodies can inhibit the cell adhesion of CGL-1 cells to MN coated plastic plates. Human anti-MN antibodies have been used to diagnose and quantify MN expression in cancer cells and tumors using FACS and immunohistochemical methods. An example is also provided where a human anti-MN IgG1 mediates tumor cell lysis though antibody-dependent cell-mediated cytotoxicity. Therefore, these antibodies will be useful for the treatment of cancers in which MN is upregulated or can be useful for the diagnosis of cancers in which MN is upregulated.

1 - 16 . (canceled) 
     
     
         17 . A nucleotide sequence that codes for a purified preparation of a human antibody, wherein the antibody binds MN protein. 
     
     
         18 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises a VH3-CDR1 region selected from the group consisting of SEQ ID NOS: 1-13. 
     
     
         19 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises a VH3-CDR3 region comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS: 14-33. 
     
     
         20 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises a VLλ1-CDR3 region comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS: 34-36. 
     
     
         21 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises a VL2-CDR1 region comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS: 37-38. 
     
     
         22 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises a VLλ2-CDR3 region comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS: 39-44. 
     
     
         23 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises VH3-CDR1, VH3-CDR3 and VL2-CDR3 nucleotide sequences selected from the group consisting of SEQ ID: 1 and 14 and 41, SEQ ID: 2 and 14 and 41, SEQ ID: 3 and 14 and 41, and SEQ ID: 4 and 14 and 41. 
     
     
         24 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises VH3-CDR3, VL2-CDR1, and VL2-CDR3 nucleotide sequences selected from the group consisting of SEQ ID: 14 and 37 and 41, and SEQ ID: 14 and 38 and 41. 
     
     
         25 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises VH3-CDR3, VL2-CDR3 and VH3-CDR1 nucleotide sequences selected from the group consisting of SEQ ID: 14 and 41 and 1, SEQ ID: 14 and 41 and 2, SEQ ID: 14 and 41 and 3, SEQ ID: 14 and 41 and 4, SEQ ID: 14 and 41 and 5, SEQ ID: 14 and 41 and 6, SEQ ID: 14 and 41 and 7, SEQ ID: 14 and 41 and 8, SEQ ID: 14 and 41 and 9, and SEQ ID: 14 and 41 and 10. 
     
     
         26 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises VH3-CDR3, VL2-CDR3 nucleotide sequences selected from the group consisting of SEQ ID: 14 and 39, SEQ ID: 15 and 42, SEQ ID: 16 and 44, SEQ ID: 17 and 39, SEQ ID: 18 and 39, SEQ ID: 19 and 40, and SEQ ID: 20 and 43. 
     
     
         27 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises VH3-CDR3, VL1-CDR3 nucleotide sequences selected from the group consisting of SEQ ID: 14 and 34, SEQ ID: 22 and 34, SEQ ID: 22 and 35, SEQ ID: 22 and 36, and SEQ ID: 23 and 34. 
     
     
         28 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises VH3-CDR3, VL2-CDR3 nucleotide sequences selected from the group consisting of SEQ ID: 14 and 41, SEQ ID: 14 and 40, SEQ ID: 14 and 42, SEQ ID: 14 and 43, SEQ ID: 14 and 44, SEQ ID: 16 and 41, SEQ ID: 16 and 40, SEQ ID: 16 and 42, SEQ ID: 16 and 43, and SEQ ID: 16 and 39. 
     
     
         29 . The purified preparation of  claim 17  wherein the nucleotide sequence comprises VH3-CDR3, VL2-CDR3 nucleotide sequences selected from the group consisting of SEQ ID: 24 and 42, SEQ ID: 14 and 42, SEQ ID: 25 and 42, SEQ ID: 26 and 42, SEQ ID: 27 and 42, SEQ ID: 28, and 42, SEQ ID: 29 and 42, SEQ ID: 30 and 42, SEQ ID: 31 and 42, SEQ ID: 32 and 42 and SEQ ID: 33 and 42. 
     
     
         30 . An expression vector comprising the polynucleotide of  claim 17 , wherein said vector codes for MN binding antibody. 
     
     
         31 . An expression vector comprising the polynucleotide of  claim 18 , wherein said vector codes for MN binding antibody. 
     
     
         32 . An expression vector comprising the polynucleotide of  claim 19 , wherein said vector codes for MN binding antibody. 
     
     
         33 . An expression vector comprising the polynucleotide of  claim 20 , wherein said vector codes for MN binding antibody. 
     
     
         34 . An expression vector comprising the polynucleotide of  claim 21 , wherein said vector codes for MN binding antibody. 
     
     
         35 . An expression vector comprising the polynucleotide of  claim 22 , wherein said vector codes for MN binding antibody. 
     
     
         36 . An expression vector comprising the polynucleotide of  claim 23 , wherein said vector codes for MN binding antibody. 
     
     
         37 . An expression vector comprising the polynucleotide of  claim 24 , wherein said vector codes for MN binding antibody. 
     
     
         38 . An expression vector comprising the polynucleotide of  claim 25 , wherein said vector codes for MN binding antibody. 
     
     
         39 . An expression vector comprising the polynucleotide of  claim 26 , wherein said vector codes for MN binding antibody. 
     
     
         40 . An expression vector comprising the polynucleotide of  claim 27 , wherein said vector codes for MN binding antibody. 
     
     
         41 . An expression vector comprising the polynucleotide of  claim 28 , wherein said vector codes for MN binding antibody. 
     
     
         42 . An expression vector comprising the polynucleotide of  claim 29 , wherein said vector codes for MN binding antibody. 
     
     
         43 . A host cell comprising the expression vector of  claim 30 , wherein said cell expresses MN binding antibody. 
     
     
         44 . A host cell comprising the expression vector of  claim 31 , wherein said cell expresses MN binding antibody. 
     
     
         45 . A host cell comprising the expression vector of  claim 32 , wherein said cell expresses MN binding antibody. 
     
     
         46 . A host cell comprising the expression vector of  claim 33 , wherein said cell expresses MN binding antibody. 
     
     
         47 . A host cell comprising the expression vector of  claim 34 , wherein said cell expresses MN binding antibody. 
     
     
         48 . A host cell comprising the expression vector of  claim 35 , wherein said cell expresses MN binding antibody. 
     
     
         49 . A host cell comprising the expression vector of  claim 36 , wherein said cell expresses MN binding antibody. 
     
     
         50 . A host cell comprising the expression vector of  claim 37 , wherein said cell expresses MN binding antibody. 
     
     
         51 . A host cell comprising the expression vector of  claim 38 , wherein said cell expresses MN binding antibody. 
     
     
         52 . A host cell comprising the expression vector of  claim 39 , wherein said cell expresses MN binding antibody. 
     
     
         53 . A host cell comprising the expression vector of  claim 40 , wherein said cell expresses MN binding antibody. 
     
     
         54 . A host cell comprising the expression vector of  claim 41 , wherein said cell expresses MN binding antibody. 
     
     
         55 . A host cell comprising the expression vector of  claim 42 , wherein said cell expresses MN binding antibody. 
     
     
         56 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 31  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         57 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 32  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         58 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 33  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         59 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 34  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         60 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 35  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         61 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 36  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         62 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 37  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         63 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 38  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         64 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 39  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         65 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 40  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         66 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 41  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         67 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 42  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         68 . A method of making a human antibody, comprising the steps of: culturing the host cell of  claim 43  under conditions whereby the antibody is expressed; and purifying the human antibody from the host cell culture. 
     
     
         69 - 87 . (canceled)