Monoclonal antibodies against human apoptosis inhibitory protein NAIP and method for assaying the NAIP

Abstract

An anti-NAIP monoclonal antibody recognizing a human apoptosis inhibitory protein NAIP having the amino acid sequence of SEQ ID NO: 1, which is produced from hybridoma prepared by fusing a myeloma cell line with antibody-producing cell of mammal immunized by antigen containing a polypeptide which comprises amino acid sequence of the 256-586th, the 841-1052nd or parts thereof in SEQ ID NO: 1, NAIP assay method using the antibody, and NAIP assay kits.

1 . An anti-NAIP monoclonal antibody, recognizing a human apoptosis inhibitory protein NAIP having the amino acid sequence of SEQ ID NO: 1, which is produced from a hybridoma prepared by fusing a myeloma cell line with an antibody-producing cell of a mammal immunized by an antigen containing a polypeptide which comprises the amino acid sequence of the 256-586th amino acids, the 841-1052nd amino acids, or parts thereof of SEQ ID NO: 1, and wherein the epitope of the anti-NAIP monoclonal antibody is in the amino acid sequence of the 256-586th amino acids, or the 841-1052nd amino acids, of SEQ ID NO: 1.  
     
     
         2 - 11 . (canceled)  
     
     
         12 . A NAIP assay kit comprising: 
 (a) a plate on which an anti-NAIP primary antibody is immobilized; and    (b) an anti-NAIP secondary antibody labeled with a marker, wherein:    (1) the primary antibody and the secondary antibody are both the anti-NAIP monoclonal antibody of  claim 1 , wherein: 
 (i) the primary antibody recognizes the amino acid sequence of the 256-585th amino acids of SEQ ID NO: 1 and the secondary antibody recognizes the amino acid sequence of the 841-1052nd amino acids of SEQ ID NO: 1; or  
 (ii) the primary antibody recognizes the amino acid sequence of the 841-1052nd amino acids of SEQ ID NO: 1 and the secondary antibody recognizes the amino acid sequence of the 256-585th amino acids of SEQ ID NO: 1; or  
   (2) the primary antibody is the anti-NAIP monoclonal antibody of  claim 1 , which recognizes the amino acid sequence of the 256-585th amino acids or the 841-1052nd amino acids of SEQ ID NO: 1 and the secondary antibody is an anti-NAIP polyclonal antibody; or    (3) the primary antibody is an anti-NAIP polyclonal antibody and the secondary antibody is the anti-NAIP monoclonal antibody of  claim 1 , which recognizes the amino acid sequence of the 256-585th amino acids or the 841-1052nd amino acids of SEQ ID NO: 1.    
     
     
         13 . (canceled)  
     
     
         14 . The NAIP assay kit of  claim 12 , wherein the marker is a radioactive isotope or a fluorescent colorant.  
     
     
         15 . The NAIP assaying kit of  claim 12 , wherein the marker is an enzyme and the kit further includes a substrate which develops a color by the enzyme activity.  
     
     
         16 . A NAIP assaying kit comprising: 
 (a) a plate on which an anti-NAIP primary antibody is immobilized;    (b) an anti-NAIP secondary antibody; and    (c) a marker to be bound with the secondary antibody, wherein:    (1) the primary antibody and the secondary antibody are both the anti-NAIP monoclonal antibody of  claim 1 , wherein: 
 (i) the primary antibody recognizes the amino acid sequence of the 256-585th amino acids of SEQ ID NO: 1 and the secondary antibody recognizes the amino acid sequence of the 841-1052nd amino acids of SEQ ID NO: 1; or  
 (ii) the primary antibody recognizes the amino acid sequence of the 841-1052nd amino acids of SEQ ID NO: 1 and the secondary antibody recognizes the amino acid sequence of the 256-585th amino acids of SEQ ID NO: 1; or  
   (2) the primary antibody is the anti-NAIP monoclonal antibody of  claim 1 , which recognizes the amino acid sequence of the 256-585th amino acids or the 841-1052nd amino acids of SEQ ID NO: 1 and the secondary antibody is an anti-NAIP polyclonal antibody; or    (3) the primary antibody is an anti-NAIP polyclonal antibody and the secondary antibody is the anti-NAIP monoclonal antibody of  claim 1 , which recognizes the amino acid sequence of the 256-585th amino acids or the 841-1052nd amino acids of SEQ ID NO: 1.    
     
     
         17 . (canceled)  
     
     
         18 . The NAIP assaying kit of  claim 16 , wherein the marker is a radioactive isotope or a fluorescent colorant.  
     
     
         19 . The NAIP assaying kit of  claim 16 , wherein the marker is an enzyme and the kit further includes a substrate which develops a color by the enzyme activity.