Novel subunits of NADH dehydrogenase

Abstract

The present invention provides four NADH dehydrogenase subunits (designated individually as NDS-1, NDS-2, NDS-3, and NDS-4 and collectively as NDS) and polynucleotides which identify and encode NDS. The invention also provides genetically engineered expression vectors and host cells comprising the nucleic acid sequences encoding NDS and a method for producing NDS. The invention also provides for use of NDS and agonists, antibodies, or antagonists specifically binding NDS, in the prevention and treatment of diseases associated with expression of NDS. Additionally, the invention provides for the use of antisense molecules to polynucleotides encoding NDS for the treatment of diseases associated with the expression of NDS. The invention also provides diagnostic assays which utilize the polynucleotide, or fragments or the complement thereof, and antibodies specifically binding NDS.

What is claimed is:  
     
         1 . An isolated antibody which specifically binds to a polypeptide comprising an amino acid sequence selected from the group consisting of: 
 a) an amino acid sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7,    b) a naturally-occurring amino acid sequence having at least 90% sequence identity to the sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7,    c) a biologically-active fragment of the amino acid sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7, and    d) an immunogenic fragment of the amino acid sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7.    
     
     
         2 . A pharmaceutical composition comprising the antibody of    claim 1    in conjunction with a suitable pharmaceutical carrier.  
     
     
         3 . A method of preparing a polyclonal antibody with the specificity of the antibody of    claim 1    comprising: 
 a) immunizing an animal with the polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7, or an antigenically-effective fragment thereof under conditions to elicit an antibody response;  
 b) isolating animal antibodies; and  
 c) screening the isolated antibodies with the polypeptide thereby identifying a polyclonal antibody binds specifically to the polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7.  
 
     
     
         4 . An antibody produced by a method of    claim 3   .  
     
     
         5 . A pharmaceutical composition comprising the antibody of    claim 4    in conjunction with a suitable pharmaceutical carrier.  
     
     
         6 . A method of making a monoclonal antibody with the specificity of the antibody of    claim 1    comprising: 
 a) immunizing an animal with the polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7, or an antigenically-effective fragment thereof under conditions to elicit an antibody response;  
 b) isolating antibody producing cells from the animal;  
 c) fusing the antibody producing cells with immortalized cells in culture to form monoclonal antibody-producing hybridoma cells;  
 d) culturing the hybridoma cells; and  
 e) isolating from the culture monoclonal antibodies which binds specifically to the polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7.  
 
     
     
         7 . A monoclonal antibody produced by a method of    claim 6   .  
     
     
         8 . A pharmaceutical composition comprising the antibody of    claim 7    in conjunction with a suitable pharmaceutical carrier.  
     
     
         9 . The antibody of    claim 1   , wherein the antibody is: 
 (a) a chimeric antibody;    (b) a single chain antibody;    (c) a Fab fragment; or    (d) a F(ab′) 2  fragment.    
     
     
         10 . The antibody of    claim 1   , wherein the antibody is produced by screening a Fab expression library.  
     
     
         11 . The antibody of    claim 1   , wherein the antibody is produced by screening a recombinant immunoglobulin library.  
     
     
         12 . A method for detecting polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7 in a sample comprising the steps of: 
 a) combining the antibody of    claim 1    with a sample under conditions to allow specific binding; and    b) detecting specific binding, wherein specific binding indicates the presence of polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7 in the sample.    
     
     
         13 . A method of using an antibody to purify polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7 from a sample, the method comprising: 
 a) combining the antibody of    claim 1    with a sample under conditions to allow specific binding; and    b) separating the antibody from the protein, thereby obtaining purified polypeptide of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:7.