C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/13 (2006.01) A61K 39/395 (2006.01) C07K 16/18 (2006.01) C07K 16/28 (2006.01) C07K 16/30 (2006.01) C12P 21/02 (2006.01) C12P 21/08 (2006.01) G01N 33/53 (2006.01) G01N 33/543 (2006.01)
Patent
CA 2463943
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.
L'invention concerne des anticorps MN monoclonaux humains ou des fragments d'anticorps MN qui ciblent la r~p~tition GEEDLP dans le domaine prot~oglycane. Le domaine prot~oglycane de la prot~ine de surface MN contient quatre de ces r~p~titions identiques GEEDLP. La liaison ~ l'~pitope souhait~ est v~rifi~e par ELISA de comp~tition, dans lequel le signal ELISA peut Útre att~nu~ par co-incubation avec un peptide contenant cette r~p~tition (PGEEDLPGEEDLP). Cette inhibition de la liaison peut ~galement Útre v~rifi~e ~ l'aide d'analyses Biacore, dans lesquelles la liaison d'anticorps souhait~s ~ MN immobilis~e ou ~ des peptides prot~oglycane peut Útre inhib~e par la r~p~tition de peptides. Outre la liaison ~ la r~p~tition de peptides, les anticorps anti-MN humains peuvent inhiber l'adh~sion cellulaire de cellules CGL-1 ~ des plaques plastiques recouverte de MN. Les anticorps anti-MN humains ont permis de diagnostiquer et de quantifier l'expression de MN dans les cellules et dans les tumeurs canc~reuses ~ l'aide de m~thodes FACS et immunohistochimiques. Dans un exemple, une anti-MN IgG1 humaine induit la lyse des cellules tumorales par une cytotoxicit~ induite par des cellules d~pendantes des anticorps. Par cons~quent, ces anticorps seront utiles pour le traitement de cancers dans lesquels MN est r~gul~ ~ la hausse ou peut Útre utile pour le diagnostic de cancers dans lesquelles MN est r~gul~ ~ la hausse.
Dubois-Stringfellow Nathalie
Murphy John E.
Rinkenberger Julie
Takeuchi Toshihiko
Bayer Healthcare Llc
Bayer Pharmaceuticals Corporation
Borden Ladner Gervais Llp
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