Target tissue-specific antigen-binding molecule

Abstract

The present inventors discovered that problems of existing antibody pharmaceuticals can be solved by producing antigen-binding molecules that contain an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a target tissue-specific compound. Use of antigen-binding molecules of the present invention enables various diseases that originate from a target tissue to be treated in a manner specific to the target tissue.

1 - 46 . (canceled) 
     
     
         47 . A method of screening for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, wherein the method comprises:
 (a) assaying binding of an antigen-binding domain to the antigen in the presence of a first concentration of a compound, wherein the compound is a small molecule natural chemical substance that is not a biopolymer, is not the antigen or a fragment thereof, and is not hydrogen ion;   (b) assaying binding of the antigen-binding domain to the antigen in the presence of a second concentration of the compound that is different from the first concentration;   (c) determining that the antigen-binding domain's binding affinity for the antigen in the presence of the first concentration of the compound is different from the antigen-binding domain's binding affinity for the antigen in the presence of the second concentration of the compound; and   (d) selecting the antigen-binding domain based on the determination of (c),   wherein either the first concentration or the second concentration, but not both, can be zero.   
     
     
         48 . The method of  claim 47 , wherein the first concentration is higher than the second concentration, and the antigen-binding domain's binding affinity for the antigen is higher in the first concentration than in the second concentration. 
     
     
         49 . The method of  claim 48 , wherein the second concentration is zero. 
     
     
         50 . A method of screening for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, wherein the method comprises:
 (a) contacting a plurality of different antigen-binding domains with an antigen in the presence of a first concentration of a compound, thereby forming one or more complexes, each comprising the antigen and an antigen-binding domain, wherein the compound is a small molecule natural chemical substance that is not a biopolymer, is not the antigen or a fragment thereof, and is not hydrogen ion;   (b) exposing the one or more complexes of (a) to a second concentration of the compound different from the first concentration, thereby causing at least one of the complexes to dissociate into its constituent antigen and antigen-binding domain; and   (c) selecting an antigen-binding domain that dissociated from the antigen in (b),   wherein either the first concentration or the second concentration, but not both, can be zero.   
     
     
         51 . The method of  claim 50 , wherein the second concentration is lower than the first concentration. 
     
     
         52 . The method of  claim 51 , wherein the second concentration is zero. 
     
     
         53 . A method of screening for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, wherein the method comprises:
 (a) contacting a plurality of different antigen-binding domains with an antigen in the presence of a first concentration of a compound, wherein the compound is a small molecule natural chemical substance that is not a biopolymer, is not the antigen or a fragment thereof, and is not hydrogen ion;   (b) isolating one or more antigen-binding domains that do not bind to the antigen in the presence of the first concentration of the compound;   (c) assaying binding of the one or more isolated antigen-binding domains to the antigen in the presence of a second concentration of the compound, wherein the second concentration is different than the first concentration, and wherein either the first concentration or the second concentration, but not both, can be zero; and   (d) selecting an antigen-binding domain that binds to the antigen in the presence of the second concentration of the compound.   
     
     
         54 . The method of  claim 53 , wherein the second concentration is higher than the first concentration. 
     
     
         55 . The method of  claim 54 , wherein the first concentration is zero. 
     
     
         56 . The method of  claim 47 , wherein the compound is selected from a purine nucleoside, a purine nucleotide, an amino acid, a metabolite of an amino acid, a lipid, a metabolite of a lipid, nicotinamide, a metabolite of nicotinamide, and a primary metabolite of glycometabolism. 
     
     
         57 . The method of  claim 47 , wherein the compound is selected from adenosine, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine, alanine, glutamic acid, aspartic acid, kynurenine, prostaglandin E2, succinic acid, citric acid, and 1-methylnicotinamide. 
     
     
         58 . The method of  claim 47 , wherein the compound is adenosine, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, or inosine. 
     
     
         59 . The method of  claim 49 , wherein step (c) comprises determining that there is at least a two-fold difference between the antigen-binding domain's binding affinity for the antigen in the presence of the first concentration of the compound and the antigen-binding domain's binding affinity for the antigen in the presence of the second concentration of the compound. 
     
     
         60 . The method of  claim 49 , wherein step (c) comprises determining that the ratio of (i) the dissociation constant (KD) of the antigen-binding domain against the antigen in the presence of the second concentration of the compound to (ii) the KD of the antigen-binding domain against the antigen in the presence of the first concentration of the compound is 2 or greater. 
     
     
         61 . The method of  claim 47 , wherein the antigen is a molecule expressed on a cell membrane. 
     
     
         62 . The method of  claim 47 , further comprising producing an antigen-binding molecule comprising the antigen-binding domain. 
     
     
         63 . The method of  claim 62 , wherein, when the antigen-binding molecule binds to the antigen, the antigen is neutralized. 
     
     
         64 . The method of  claim 62 , wherein the antigen-binding molecule has cytotoxic activity. 
     
     
         65 . The method of  claim 47 , wherein the antigen-binding domain comprises an antibody heavy-chain variable region and an antibody light-chain variable region. 
     
     
         66 . The method according to  claim 62 , wherein the antigen-binding molecule is an antibody or antibody fragment. 
     
     
         67 . The method of  claim 62 , wherein the antigen-binding molecule comprises an Fc region. 
     
     
         68 . The method of  claim 62 , wherein the method comprises:
 (i) providing a cell containing DNA encoding the antigen-binding molecule;   (ii) expressing the DNA, thereby producing the antigen-binding molecule; and   (iii) collecting the antigen-binding molecule.   
     
     
         69 . The method of  claim 50 , wherein the compound is selected from a purine nucleoside, a purine nucleotide, an amino acid, a metabolite of an amino acid, a lipid, a metabolite of a lipid, nicotinamide, a metabolite of nicotinamide, and a primary metabolite of glycometabolism. 
     
     
         70 . The method of  claim 50 , wherein the compound is selected from adenosine, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine, alanine, glutamic acid, aspartic acid, kynurenine, prostaglandin E2, succinic acid, citric acid, and 1-methylnicotinamide. 
     
     
         71 . The method of  claim 50 , wherein the compound is adenosine, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, or inosine. 
     
     
         72 . The method of  claim 50 , wherein the antigen is a molecule expressed on a cell membrane. 
     
     
         73 . The method of  claim 50 , further comprising producing an antigen-binding molecule comprising the selected antigen-binding domain. 
     
     
         74 . The method of  claim 73 , wherein, when the antigen-binding molecule binds to the antigen, the antigen is neutralized. 
     
     
         75 . The method of  claim 73 , wherein the antigen-binding molecule has cytotoxic activity. 
     
     
         76 . The method of  claim 50 , wherein the selected antigen-binding domain comprises an antibody heavy-chain variable region and an antibody light-chain variable region. 
     
     
         77 . The method according to  claim 73 , wherein the antigen-binding molecule is an antibody or antibody fragment. 
     
     
         78 . The method of  claim 73 , wherein the antigen-binding molecule comprises an Fc region. 
     
     
         79 . The method of  claim 73 , wherein the method comprises:
 (i) providing a cell containing DNA encoding the antigen-binding molecule;   (ii) expressing the DNA, thereby producing the antigen-binding molecule; and   (iii) collecting the antigen-binding molecule.   
     
     
         80 . The method of  claim 53 , wherein the compound is selected from a purine nucleoside, a purine nucleotide, an amino acid, a metabolite of an amino acid, a lipid, a metabolite of a lipid, nicotinamide, a metabolite of nicotinamide, and a primary metabolite of glycometabolism. 
     
     
         81 . The method of  claim 53 , wherein the compound is selected from adenosine, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine, alanine, glutamic acid, aspartic acid, kynurenine, prostaglandin E2, succinic acid, citric acid, and 1-methylnicotinamide. 
     
     
         82 . The method of  claim 53 , wherein the compound is adenosine, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, or inosine. 
     
     
         83 . The method of  claim 53 , wherein the antigen is a molecule expressed on a cell membrane. 
     
     
         84 . The method of  claim 53 , further comprising producing an antigen-binding molecule comprising the selected antigen-binding domain. 
     
     
         85 . The method of  claim 84 , wherein, when the antigen-binding molecule binds to the antigen, the antigen is neutralized. 
     
     
         86 . The method of  claim 84 , wherein the antigen-binding molecule has cytotoxic activity. 
     
     
         87 . The method of  claim 53 , wherein the selected antigen-binding domain comprises an antibody heavy-chain variable region and an antibody light-chain variable region. 
     
     
         88 . The method according to  claim 84 , wherein the antigen-binding molecule is an antibody or antibody fragment. 
     
     
         89 . The method of  claim 84 , wherein the antigen-binding molecule comprises an Fc region. 
     
     
         90 . The method of  claim 84 , wherein the method comprises:
 (i) providing a cell containing DNA encoding the antigen-binding molecule;   (ii) expressing the DNA, thereby producing the antigen-binding molecule; and   (iii) collecting the antigen-binding molecule.