Antibodies to crosslinkers and methods for using the same

Abstract

The present invention is directed to the discovery of antibodies that will react to proteins or nucleic acids bound to particular crosslinkers, but not to the free crosslinkers or free proteins or nucleic acids. Monoclonal antibodies with such binding specificity have widespread applications in receptor-ligand binding, immunodiagnostic and nucleic acid diagnostics.

We claim:  
     
         1 . A monoclonal antibody that will specifically bind to a BS3 or DSS modified protein or nucleic acid, but not to free BS3 or DSS or unmodified protein or nucleic acid.  
     
     
         2 . The monoclonal antibody of  claim 1 , which is selected from the group consisting of: 7E3-2E7; 2C3-2E10; 12G9-2C5; and 11F2-2F7.  
     
     
         3 . A nucleic acid molecule which has been modified by covalent bonding to BS3 or DSS.  
     
     
         4 . The nucleic acid molecule of  claim 3 , which is a hybridization probe in a diagnostic assay.  
     
     
         5 . A method of assaying receptor-ligand binding, comprising: 
 (a) linking BS3 or DSS to the ligand of interest;    (b) reacting the BS3 or DSS modified ligand with cells expressing the ligand's receptor;    (c) reacting bound BS3 or DSS modified ligand/receptor with labeled anti-BS3/DSS monoclonal antibody; and    (d) detecting the label on a complex formed between the labeled antibody and the receptor/ligand.    
     
     
         6 . A method of assaying receptor-ligand binding, comprising: 
 (a) linking BS3 or DSS to the ligand of interest;    (b) reacting the BS3 or DSS modified ligand with cells expressing the ligand's receptor;    (c) reacting bound BS3 or DSS modified ligand/receptor with anti-BS3/DSS monoclonal antibody, followed by anti-mouse IgG conjugated to FITC; and    (d) detecting the bound FITC-conjugated antibody by flow cytometry or immunofluorescence.    
     
     
         7 . The method of  claim 6 , wherein the ligand is gp120 and the receptor is CD4.  
     
     
         8 . A method for detecting products of a nucleic acid amplification, comprising: 
 (a) labelling NTPs that contain a free NH 2  group with BS3 or DSS;    (b) using the labelled NTPs in an amplification reaction mixture;    (c) at the conclusion of the amplification, reacting the amplification products with anti-BS3/DSS antibody, which is itself labelled with a detectable moiety or reacted with a labelled antibody that will bind to the anti-BS3/DSS antibody, thereby detecting the amplification products.    
     
     
         9 . The method of  claim 8 , wherein the amplification is an isothermal transcription based amplification reaction and the NTPs are rNTPs.  
     
     
         10 . The method of  claim 9 , wherein the amplification reaction is NASBA.  
     
     
         11 . A method for detecting a nucleic acid sequence, comprising hybridizing a probe with complementary regions of said sequence, wherein said probe comprises a nucleic acid sequence labeled with BS3 or DSS.  
     
     
         12 . An immunoassay using a labeled antigen or a labeled antibody, wherein BS3 or DSS is the label.  
     
     
         13 . A method for detecting an antigen of interest in a mixture of antigens, comprising: 
 (a) modifying the antigens in the sample with BS3 or an analogue thereof;    (b) reacting the modified antigens with a capture antibody, which is an IgA anti-BS3 antibody;    (c) reacting the bound antigen with an IgG antibody that is specific to the antigen of interest; and    (d) detecting bound IgG by reacting with a labeled anti-IgG antibody,    thereby detecting the antigen of interest.    
     
     
         14 . The method of  claim 13 , wherein the IgA antibody is 7E3-2E7.