Antibodies to oxidized phospholipids

Abstract

The disclosure provides for single chain variable fragments to oxidized phospholipid epitopes and methods of use thereof, including the production of transgenic animal models and the use of the fragments as therapeutic agents for treating CAS.

1 . A chimeric or humanized antibody or antibody fragment that recognizes and binds to a phosphocholine headgroup of an oxidized phospholipid, wherein the chimeric or humanized antibody or antibody fragment comprises a variable heavy chain (V H ) domain and/or a variable light chain (V L ) domain, and wherein (a) the V H  domain comprises an amino acid sequence that includes one, two or three complementarity determining regions (CDRs) selected from the group consisting of: SEQ ID NO:6 and sequence that are at least 95%, 96%, 97%, 98%, 99% or 99.9% identical to SEQ ID NO:6; SEQ ID NO:7 and sequence that are at least 95%, 96%, 97%, 98%, 99% or 99.9% identical to SEQ ID NO:7; and SEQ ID NO:8 and sequence that are at least 95%, 96%, 97%, 98%, 99% or 99.9% identical to SEQ ID NO:8; and (b) the V L  domain comprises an amino acid sequence that includes one, two or three complementarity determining regions (CDRs) selected from the group consisting of: SEQ ID NO:9 or 12 and sequence that are at least 95%, 96%, 97%, 98%, 99% or 99.9% identical to SEQ ID NO:9 or 12; SEQ ID NO:10 and sequence that are at least 95%, 96%, 97%, 98%, 99% or 99.9% identical to SEQ ID NO:10; and SEQ ID NO:11 and sequence that are at least 95%, 96%, 97%, 98%, 99% or 99.9% identical to SEQ ID NO:11. 
     
     
         2 . The chimeric or humanized antibody or antibody fragment of  claim 1 , wherein the V H  domain comprises an amino acid sequence that includes CDRs comprising SEQ ID NO:6, 7 and 8, and/or the V L  domain comprises an amino acid sequence that includes CDRs comprising SEQ ID NO:9, 10 and 11, or SEQ ID NO:10, 11 and 12. 
     
     
         3 . The antibody or an antibody fragment of  claim 1 , wherein the antibody or antibody fragment is selected from the group consisting of: (a) an antibody or scFv with heavy and light chain domains comprising the complementarity determining regions of SEQ ID NO:6, 7, 8, 9, 10 and 11; and (b) an antibody or scFv with heavy and light chain domains comprising the complementarity determining regions of SEQ ID NO:6, 7, 8, 10, 11 and 12. 
     
     
         4 . The chimeric or humanized antibody or antibody fragment of  claim 1 , wherein the heavy and light chain domains are linked to an Fc region. 
     
     
         5 . A single chain variable fragment (“scFv”), wherein the scFv is the antibody fragment of  claim 1  wherein the scFv recognizes a phosphocholine headgroup of an oxidized phospholipid. 
     
     
         6 . The scFv of  claim 5 , wherein the scFv is soluble under physiological conditions. 
     
     
         7 . The scFv of  claim 5 , wherein the scFv is murine. 
     
     
         8 . The scFv of  claim 5 , wherein the scFv is humanized. 
     
     
         9 . The scFv of  claim 5 , wherein the scFv comprises a light-chain variable region having a sequence that is at least 95% identical to the sequence as set forth in SEQ ID NO:2 from amino acid 34 to about amino acid 146. 
     
     
         10 . The scFv of  claim 9 , wherein the scFv comprises a heavy chain variable region having a sequence that is at least 95% identical to the sequence as set forth in SEQ ID NO:2 from about amino acid 162 to about amino acid 284. 
     
     
         11 . The scFv of  claim 5 , wherein the scFv comprises a light-chain variable region having a sequence that is at least 95% identical to the sequence as set forth in SEQ ID NO:4 from amino acid 24 to about amino acid 135. 
     
     
         12 . The scFv of  claim 11 , wherein the scFv comprises a heavy chain variable region having a sequence that is at least 95% identical to the sequence as set forth in SEQ ID NO:4 from about amino acid 151 to about amino acid 273. 
     
     
         13 . The scFv of  claim 5 , wherein the scFv comprises a sequence that is at least 95% identical to the sequence as set forth in SEQ ID NO:2. 
     
     
         14 . The scFv of  claim 5 , wherein the scFv comprises a sequence that is at least 95% identical to the sequence as set forth in SEQ ID NO:4 from amino acid 1 to about amino acid 273. 
     
     
         15 - 16 . (canceled) 
     
     
         17 . The chimeric antibody of  claim 1 , wherein the variable light chain and variable heavy chain are non-human and an Fc region is human or humanized. 
     
     
         18 . The chimeric antibody of  claim 1 , wherein the variable light chain and variable heavy chain are human or humanized and the Fc region is non-human. 
     
     
         19 . (canceled) 
     
     
         20 . The humanized antibody of  claim 1 , wherein the antibody comprises a sequence that is at least 95% identical to a sequence as set forth in SEQ ID NO:4. 
     
     
         21 . A polynucleotide that encodes an antibody, antibody fragment of  claim 1 . 
     
     
         22 . The polynucleotide of  claim 21 , wherein the polynucleotide comprises a sequence that is at least 95% identical to SEQ ID NO:1 or 3 and encodes a polypeptide that has the binding specificity of an E06 antibody. 
     
     
         23 . A vector comprising a polynucleotide of  claim 21 . 
     
     
         24 . A host cell transformed with the polynucleotide of  claim 21 . 
     
     
         25 . A host cell transformed with a vector of  claim 23 . 
     
     
         26 . A transgenic animal, comprising a polynucleotide of  claim 21 . 
     
     
         27 . The transgenic animal of  claim 26 , wherein the animal is a mouse. 
     
     
         28 . The transgenic animal of  claim 26 , wherein the polynucleotide is expressed from hepatocytes and/or macrophages. 
     
     
         29 . (canceled) 
     
     
         30 . A method of treating a subject with an inflammatory disease, disorder or condition or a disease or condition associated with Oxidized phospholipids by administering a chimeric or humanized antibody or antibody fragment of  claim 1 . 
     
     
         31 . A method of treating or preventing calcific aortic stenosis (CAS) comprising administering a chimeric or humanized antibody or antibody fragment of  claim 1  to a subject having or at risk of having CAS.