Antibodies to TTR and Methods of Use

Abstract

The disclosure pertains to antibodies and binding fragments thereof that specifically binds all or part of EHAEVVFTA. Also provided are isolated peptides, isolated nucleic acids, immunogens, compositions, immunoassays and kits and method of using said reagents to detect misfolded TTR.

1 . An antibody or binding fragment thereof that specifically binds all or part of EHAEVVFTA, wherein the part comprises at least 4 or at least 5 or more contiguous amino acids of EHAEVVFTA. 
     
     
         2 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody or binding fragment specifically binds misfolded and/or monomeric TTR, optionally TTR fibrils. 
     
     
         3 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody or binding fragment thereof is capable of disrupting or reducing TTR fibril formation when monomeric TTR or misfolded TTR intermediates in solution is/are contacted with the antibody or binding fragment. 
     
     
         4 . The antibody or binding fragment thereof of  claim 1 , wherein the antibody is a monoclonal or polyclonal antibody and/or a chimeric and/or a humanized antibody or binding fragment thereof. 
     
     
         5 . The antibody binding fragment of  claim 1 , wherein the antibody fragment is a Fab, Fab′, F(ab′) 2 , scFv, dsFv, ds-scFv, dimer, minibody, diabody, or multimer thereof or a bispecific antibody fragment. 
     
     
         6 . The antibody of  claim 1 , wherein the antibody is affinity purified. 
     
     
         7 . An isolated peptide comprising all or part of EHAEVVFTA, the part comprising at least 4 or at least 5 or more contiguous amino acids. 
     
     
         8 . The isolated peptide of  claim 7 , further comprising one or more amino acid linkers conjugated to one or more of the peptide N and/or C termini, optionally wherein the amino acid linkers include one or more G or K or S residues and for example the amino acid linker or linkers each independently comprise GG or GGKG or glycine serine linkers. 
     
     
         9 . The isolated peptide of  claim 7  comprising a peptide comprising all or part of EHAEVVFTA and a tag. 
     
     
         10 . An isolated nucleic acid encoding the isolated peptide of a  claim 7 . 
     
     
         11 . An immunogen comprising the peptide of  claim 7 , optionally further comprising an immunogenicity enhancing molecule conjugated to the peptide. 
     
     
         12 . A composition comprising:
 a. the antibody of  claim 1 , and optionally a suitable carrier: or   b. an immunogen comprising a peptide comprising all or part of EHAEVVFTA,   the part comprising at least 4 or at least 5 or more contiguous amino acids of EHAEVVFTA and optionally an adjuvant, optionally selected from Freund's complete and incomplete adjuvant.   
     
     
         13 . An immunoassay comprising the antibody of  claim 1 , optionally an ELISA. 
     
     
         14 . A kit comprising:
 a. the antibody of  claim 1 ;   b. an isolated peptide comprising all or part of EHAEVVFTA, the part comprising at least 4 or at least 5 or more contiguous amino acids,   c. an isolated nucleic acid encoding the isolated peptide of (b),   d. an immunogen comprising the isolated peptide of (b),   e. a composition comprising (a), (b), (c) or (d), and/or   f. an immunoassay comprising the antibody of (a), and   g. a compartment such as a vial for housing the antibody, isolated peptide, isolated nucleic acid, immunogen and/or composition, and   h. optionally instructions for use in a method described herein.   
     
     
         15 . A method for preparing an antibody, the method comprising immunizing a non-human subject with the immunogen of  claim 11  or a composition comprising said immunogen and optionally isolating an antibody that specifically binds EHAEVVFTA. 
     
     
         16 . The method of  claim 15  further comprising forming an antibody-producing hybridoma. 
     
     
         17 . A method of detecting a misfolded TTR conformation and/or monomeric TTR in a test sample, the method comprising contacting the test sample with an antibody or binding fragment thereof of  claim 1  under conditions suitable to form a specific antibody antigen complex between the antibody and TTR; determining if a specific antibody TTR complex is formed thereby detecting misfolded TTR and/or monomeric TTR in the test sample. 
     
     
         18 . The method of  claim 17 , wherein the determining comprises measuring the amount of the antibody-antigen complex in the test sample and comparing the amount of antibody-antigen complex in the test sample to a control, wherein a difference in the amount of antibody-antigen complex in the test sample as compared to the control is indicative that misfolded TTR is detected in the test sample. 
     
     
         19 . The method of  claim 17  for detecting a TTR amyloid deposition disease, optionally wherein the TTR amyloid deposition disease is selected from Senile systemic amyloidosis, familial amyloid polyneuropathy, familial amyloid cardiomyopathy. 
     
     
         20 . The method of  claim 19 , wherein the test sample is a biological sample obtained from a subject, optionally wherein the sample comprises blood, serum, plasma and/or solid tissue. 
     
     
         21 . A method of inhibiting TTR fibril formation, the method comprising contacting an antibody or fragment of  claim 1  with misfolded TTR or monomeric TTR in solution under fibril formation conditions, wherein the antibody specifically binds the misfolded and/or monomeric TTR and inhibits fibril formation, optionally wherein the antibody or binding fragment is contracted with misfolded TTR or monomeric TTR during the lag phase of fibril formation.