Antibodies recognizing medin

Abstract

The invention provides antibodies that specifically bind to medin. The antibodies have the capacity to bind to monomeric, misfolded, aggregated, fibril or deposited forms of medin. The antibodies can be used for treating or effecting prophylaxis of diseases associated with medin, medin accumulation or accumulation of medin deposits (e.g., medin amyloidosis). The antibodies can also be used for diagnosing medin amyloidosis and inhibiting or reducing aggregation of medin, among other applications.

1 - 113 . (canceled) 
     
     
         114 . A method of identifying an antibody that binds to an epitope within amino acid residues 44-50 of SEQ ID NO:1, comprising:
 (a) immunizing a non-human animal with human medin or a fragment thereof; and   (b) screening induced antibodies to identify an antibody binding within amino acid residues 44-50 of SEQ ID NO:1.   
     
     
         115 . The method of  claim 114 , wherein the fragment consists of amino acid residues 44-50 of SEQ ID NO:1, optionally linked to a carrier. 
     
     
         116 . The method of  claim 114 , wherein the screening is performed by determining binding of the antibodies to a fragment of human medin consisting of residues 44-50 of SEQ ID NO:1. 
     
     
         117 . A method of identifying an antibody comprising:
 (a) immunizing a non-human animal with human medin or a fragment thereof to induce antibodies;   (b) screening the induced antibodies to identify an antibody that specifically binds medin or a fragment thereof and does not specifically bind to native lactadherin.   
     
     
         118 . The method of  claim 117 , wherein the fragment consists of amino acid residues 44-50 of SEQ ID NO:1, optionally linked to a carrier. 
     
     
         119 . A method of generating an antibody that competes with a reference antibody for binding to human medin comprising immunizing an animal or B cells with an immunogen comprising human medin or a fragment thereof to produce antibodies, performing a competition assay with the produced antibodies, the reference antibody, and human medin to identify a subset of produced antibodies that inhibit binding of the reference antibody to the human medin; wherein the reference antibody is an antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:11 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:29 or is an antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:3 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:36. 
     
     
         120 . The method of  claim 119 , wherein the reference antibody is an antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:11 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:29. 
     
     
         121 . The method of  claim 120 , wherein the fragment consists of amino acid residues 44-50 of SEQ ID NO:1, optionally linked to a carrier. 
     
     
         122 . The method of  claim 120 , wherein the reference antibody is an antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:3 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:36. 
     
     
         123 . A method of producing an antibody that binds the same epitope on human medin as a reference antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:3 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:36, the method comprising:
 (a) immunizing a non-human animal with medin or a fragment thereof;   (b) screening induced antibodies for differential binding to wild-type medin or fragments thereof compared to mutagenized forms of the medin antigen; and   (c) selecting an antibody, wherein the same mutagenized forms significantly reduce the binding of both the induced antibody and reference antibody to medin or a fragment thereof.   
     
     
         124 . A method of producing an antibody that binds the same epitope on human medin as a reference antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:11 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:29, the method comprising:
 (a) immunizing a non-human animal with medin or a fragment thereof;   (b) screening induced antibodies for differential binding to wild-type medin or fragments thereof compared to mutagenized forms of the medin antigen; and   (c) selecting an induced antibody, wherein the same mutagenized forms significantly reduce the binding of both the induced antibody and the reference antibody to medin or a fragment thereof.   
     
     
         125 . The method of  claim 124 , wherein the mutagenized forms of the medin antigen comprise single amino acid replacements of a wild-type medin amino acid residue with alanine. 
     
     
         126 . The method of  claim 124 , wherein the mutagenized forms of the medin antigen comprise single amino acid replacements of a wild-type medin amino acid residue with serine. 
     
     
         127 . A method of  claim 124 , further comprising producing a humanized form of the antibody, the method comprising:
 (a) synthesizing a nucleic acid encoding a humanized heavy chain comprising CDRs of the non-human animal antibody heavy chain and a nucleic acid encoding a humanized light chain comprising CDRs of the non-human animal antibody light chain; and   (b) expressing the nucleic acids in a host cell to produce a humanized antibody.