Compositions and methods for binding lysophosphatidic acid

Abstract

Compositions and methods for making and using anti-LPA agents, for example, monoclonal antibodies, are described.

What is claimed is: 
     
         1 . A method of detecting LPA or a metabolite thereof in a sample obtained from a subject, comprising detecting binding of LPA or a metabolite thereof in a sample to an antibody, or antigen-binding fragment thereof, that specifically binds LPA or a metabolite thereof, under conditions that allow the antibody or antigen-binding fragment thereof to bind to the LPA or metabolite thereof if present in the sample, wherein the antibody or antigen-binding fragment thereof comprises at least one immunoglobulin heavy chain variable domain and at least one immunoglobulin light chain variable domain, wherein:
 (i) each immunoglobulin heavy chain variable domain comprises first, second, and third heavy chain complementarity determining regions (CDRs), wherein the first heavy chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 56, 68, and 91, the second heavy chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 57, 69, 79, and 92, and the third heavy chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 58, 70, 80, 84, and 93; and   (ii) each immunoglobulin light chain variable domain comprises first, second, and third light chain CDRs, wherein the first light chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 59, 71, 81, and 94, the second light chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO; 60, 72, 82, and 95, and the third light chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 61 and 96.   
     
     
         2 . A method according to  claim 1  wherein the sample is an animal-derived sample, and optionally wherein the subject is a mammal, optionally a human. 
     
     
         3 . A method according to  claim 2  wherein the animal-derived sample is selected from the group consisting of a tissue sample and a bodily fluid sample. 
     
     
         4 . A method according to  claim 3  wherein the tissue sample is a biopsy sample. 
     
     
         5 . A method according to  claim 3  wherein the bodily fluid sample is selected from the group consisting of whole blood, plasma, serum, urine, semen, bile, aqueous humor, vitreous humor, mucus, bronchioalveolar lavage fluid, and sputum. 
     
     
         6 . A method according to  claim 1  wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof. 
     
     
         7 . A method according to  claim 1  further comprising measuring an amount of LPA or metabolite thereof in the sample. 
     
     
         8 . A method according to  claim 7  wherein the method further comprises comparing a level of LPA or metabolite thereof in the sample to a reference level of LPA or metabolite thereof obtained from a normal animal of the same species as the subject, wherein the presence of an increased level of LPA or metabolite thereof relative to the reference level correlates with the presence of disease. 
     
     
         9 . A method according to  claim 7  wherein the method further comprises comparing a level of LPA or metabolite thereof in the sample to a desired level of LPA or metabolite thereof, and, if necessary, altering a therapeutic dosage of an anti-LPA agent administered to the subject, wherein the anti-LPA agent modulates the effective concentration of LPA, in order to regulate the effective concentration of LPA in the subject. 
     
     
         10 . A diagnostic kit for detecting lysophosphatidic acid (LPA) for use in a method according to  claim 1 , comprising:
 (a) a diagnostic reagent comprising a derivatized LPA that comprises a hydrocarbon chain, wherein a carbon atom within the hydrocarbon chain is derivatized with a reactive group; and   (b) an antibody, or antigen-binding fragment thereof, that specifically binds LPA, wherein the antibody or antigen-binding fragment thereof comprises at least one immunoglobulin heavy chain variable domain and at least one immunoglobulin light chain variable domain, wherein:
 (i) each immunoglobulin heavy chain variable domain comprises first, second, and third heavy chain complementarity determining regions (CDRs), wherein the first heavy chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 56, 68, and 91, the second heavy chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 57, 69, 79, and 92, and the third heavy chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 58, 70, 80, 84, and 93; and 
 (ii) each immunoglobulin light chain variable domain comprises first, second, and third light chain CDRs, wherein the first light chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 59, 71, 81, and 94, the second light chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO; 60, 72, 82, and 95, and the third light chain CDR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 61 and 96. 
   
     
     
         11 . A diagnostic kit according to  claim 10  wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof. 
     
     
         12 . A diagnostic kit according to  claim 10  wherein the reactive group is selected from the group consisting of a sulfhydryl (thiol) group, a carboxylic acid group, a cyano group, an ester, a hydroxy group, an alkene, an alkyne, an acid chloride group, and a halogen atom. 
     
     
         13 . A diagnostic kit according to  claim 10  wherein the derivatized LPA is associated with a solid support. 
     
     
         14 . A diagnostic kit according to  claim 13  wherein the derivatized LPA is covalently associated with the solid support. 
     
     
         15 . A diagnostic kit according to  claim 10  wherein the derivatized LPA is conjugated to a carrier moiety selected from the group consisting of polyethylene glycol, colloidal gold, adjuvant, a silicone bead, and a protein. 
     
     
         16 . A diagnostic kit according to  claim 15  wherein the protein is selected from the group consisting of keyhole limpet hemocyanin, albumin, bovine thyroglobulin, and soybean trypsin inhibitor. 
     
     
         17 . A diagnostic kit according to  claim 10  wherein the diagnostic reagent is a thiolated LPA conjugated to bovine serum albumin or keyhole limpet hemocyanin. 
     
     
         18 . A diagnostic kit according to  claim 15  wherein the carrier moiety is associated with a solid support. 
     
     
         19 . A diagnostic kit according to  claim 10  that is an ELISA kit.