Hyaluronan receptor for endocytosis

Abstract

A purified nucleic acid segment encoding a functionally active hyaluronan receptor for endocytosis (HARE) or an active peptide fragment thereof, and methods for producing functionally active HARE or an active peptide fragment thereof therefrom, wherein the functionally active HARE or an active peptide fragment thereof is able to specifically bind HA, chondroitin and chondroitin sulfate.

1 . A purified antibody that binds specifically to a HARE protein.  
     
     
         2 . The purified antibody of  claim 1 , wherein the HARE protein is selected from the group consisting of: 
 (a) a HARE protein in accordance with SEQ ID NO:2;    (b) a HARE protein in accordance with SEQ ID NO:25;    (c) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:1;    (d) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:24; and    (e) a HARE protein having at least 78% identity to the HARE protein in accordance with SEQ ID NO:25.    
     
     
         3 . The purified antibody of  claim 1 , wherein the purified antibody specifically inhibits binding of at least one of HA, chondroitin and chondroitin sulfate to HARE.  
     
     
         4 . A purified antibody that selectively binds to an epitope of a HARE protein.  
     
     
         5 . The purified antibody of  claim 4 , wherein the epitope is in an HA-binding domain of the HARE protein.  
     
     
         6 . The purified antibody of  claim 4 , wherein the purified antibody inhibits the binding of at least one of HA, chondroitin and chondroitin sulfate to HARE.  
     
     
         7 . The purified antibody of  claim 4 , wherein the HARE protein is selected from the group consisting of: 
 (a) a HARE protein in accordance with SEQ ID NO:2;    (b) a HARE protein in accordance with SEQ ID NO:25;    (c) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:1;    (d) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:24; and    (e) a HARE protein having at least 78% identity to the HARE protein in accordance with SEQ ID NO:25.    
     
     
         8 . A purified antibody that binds specifically to a HARE protein and inhibits the binding of at least one of HA, chondroitin and chondroitin sulfate to the HARE protein.  
     
     
         9 . A monoclonal antibody having the same epitope specificity as at least one of: 
 (a) mAb-28, ATCC accession number PTA-5354;    (b) mAb-30, ATCC accession number PTA-5355;    (c) mAb-54, ATCC accession number PTA-5356;    (d) mAb-159, ATCC accession number PTA-5358;    (e) mAb-174, ATCC accession number PTA-5359;    (f) mAb-235, ATCC accession number PTA-5359; and    (g) mAb-467, ATCC accession number PTA-5361.    
     
     
         10 . A method of making an antibody, comprising immunizing a non-human animal with an immunogenic fragment of a HARE protein.  
     
     
         11 . The method of  claim 10  wherein the immunogenic fragment is selected from the group consisting of: 
 (a) SEQ ID NO:2;    (b) SEQ ID NO:25;    (c) an amino acid sequence encoded by SEQ ID NO:1;    (d) an amino acid sequence encoded by SEQ ID NO:24;    (e) an amino acid sequence having at least 78% identity to SEQ ID NO:25; and    (f) at least a portion of any of (a)-(e).    
     
     
         12 . The method of  claim 10  wherein the immunogenic fragment is an HA-binding domain of a HARE protein.  
     
     
         13 . A method of making a humanized monoclonal antibody, comprising: 
 immunizing a non-human animal with an immunogenic fragment of a HARE protein to form a monoclonal antibody; and    removing portions of the monoclonal antibody which may be antigenic in a human and substituting like portions of a human antibody therefor.    
     
     
         14 . The method of  claim 13  wherein the immunogenic fragment is selected from the group consisting of: 
 (a) SEQ ID NO:2;    (b) SEQ ID NO:25;    (c) an amino acid sequence encoded by SEQ ID NO:1;    (d) an amino acid sequence encoded by SEQ ID NO:24;    (e) an amino acid sequence having at least 78% identity to SEQ ID NO:25; and    (f) at least a portion of any of (a)-(e).    
     
     
         15 . The method of  claim 14  wherein the immunogenic fragment comprises an HA-binding domain of a HARE protein.  
     
     
         16 . A method of making a monoclonal antibody, comprising the steps of: 
 identifying a hybridoma cell that produces a monoclonal antibody specific for a HARE protein; and    culturing the hybridoma cell under conditions that permit production of the monoclonal antibody.    
     
     
         17 . The method of  claim 16  wherein, in the step of identifying a hybridoma cell, the HARE protein is selected from the group consisting of: 
 (a) a HARE protein in accordance with SEQ ID NO:2;    (b) a HARE protein in accordance with SEQ ID NO:25;    (c) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:1;    (d) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:24; and    (e) a HARE protein having at least 78% identity to the HARE protein in accordance with SEQ ID NO:25.    
     
     
         18 . The method of  claim 16  wherein the method further comprises isolating the monoclonal antibody from the cultured hybridoma cell.  
     
     
         19 . A method of purifying a functionally active HARE protein wherein the HARE protein is able to specifically bind at least one of HA, chondroitin and chondroitin sulfate, comprising the steps of: 
 providing a biological sample containing a HARE protein;    providing an affinity matrix comprising: 
 an antibody that selectively binds to an epitope of a HARE protein; and  
 a solid support to which the antibody is bound;  
   contacting the biological sample with the affinity matrix such that the HARE protein in the biological sample binds to the antibody of the affinity matrix;    separating the HARE protein bound to the antibody of the affinity matrix from the remainder of the biological sample; and    releasing the HARE protein from the antibody of the affinity matrix.    
     
     
         20 . The method of  claim 19  wherein, in the step of providing the biological sample, the biological sample is obtained from at least one of liver, spleen, lymph nodes and bone marrow.  
     
     
         21 . The method of  claim 19  wherein, in the step of providing an affinity matrix, the antibody of the affinity matrix inhibits the binding of at least one of HA, chondroitin and chondroitin sulfate to the HARE protein.  
     
     
         22 . The method of  claim 19  wherein the step of releasing the HARE protein from the antibody of the affinity matrix is further defined as contacting the HARE protein bound to the antibody of the affinity matrix with an eluate which disrupts the binding between the HARE protein and the antibody.  
     
     
         23 . The method of  claim 19  wherein, in the step of providing an affinity matrix, the antibody of the affinity matrix is a monoclonal antibody.  
     
     
         24 . The method of  claim 19  wherein, in the step of providing an affinity matrix comprising an antibody that selectively binds to an epitope of a HARE protein, the HARE protein is selected from the group consisting of: 
 (a) a HARE protein in accordance with SEQ ID NO:2;    (b) a HARE protein in accordance with SEQ ID NO:25;    (c) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:1;    (d) a HARE protein encoded by the nucleic acid sequence of SEQ ID NO:24; and    (e) a HARE protein having at least 78% identity to the HARE protein in accordance with SEQ ID NO:25.