Antibody specific capture agents, compositions, and methods of using and making

Abstract

The present application provides stable peptide-based antibody capture agents and methods of use as detection and diagnosis agents. The application further provides methods of manufacturing antibody capture agents using iterative on-bead in situ click chemistry.

1 . A stable, synthetic capture agent that specifically binds anti-gp41 antibodies, wherein the capture agent comprises a designed anchor ligand, a designed secondary ligand, optionally, a designed tertiary ligand, and optionally a designed quarternary ligand and wherein the ligands selectively bind anti-gp41 antibodies. 
     
     
         2 . The capture agent of  claim 1 , wherein the anchor ligand comprises an amino acid sequence 80 to 100% identical to IWCGSGKLICTTA (SEQ ID NO:1). 
     
     
         3 . The capture agent of  claim 2 , wherein the anchor ligand comprises the amino acid sequence of IWCGSGKLICTTA (SEQ ID NO:1). 
     
     
         4 . The capture agent of  claim 2 , wherein the secondary ligand comprises an amino acid sequence 80 to 100% identical to an amino acid sequence selected from the group consisting of dkGlp (SEQ ID NO: 8); vadpa (SEQ ID NO: 9); nidnG (SEG ID NO:2); pGvtf (SEQ ID NO:10); srrs (SEQ ID NO: 11); dq/eGaf (SEQ ID NO: 12); ywdyn (SEQ ID NO. 13); hllyq (SEQ ID NO: 14); sGaqs (SEQ ID NO:15); ddwai (SEQ ID NO: 16); qidlr (SEQ ID NO: 17); tfLqs (SEQ ID NO: 18); hnpfk (SEQ ID NO. 3); wGehp (SEQ ID NO: 19); qndwK (SEQ ID NO: 20); ltsrY (SEQ ID NO: 21); diksp (SEQ ID NO: 22); eihny (SEQ ID NO. 4); qpidq (SEQ ID NO. 23); kidrv (SEQ ID NO. 24); qsp/dwl (SEQ ID NO: 25); and eqtfd (SEQ ID NO: 26). 
     
     
         5 . The capture agent of  claim 4 , wherein the secondary ligand comprises an amino acid sequence 80 to 100% identical to an amino acid sequence selected from the group consisting of nidnG (SEQ ID NO:2), hnpfk (SEQ ID NO:3) and eihny (SEQ ID NO:4). 
     
     
         6 . The capture agent of  claim 5 , wherein the secondary ligand comprises the amino acid sequence of nidnG (SEQ ID NO:2). 
     
     
         7 . The capture agent of  claim 5 , wherein the secondary ligand comprises the amino acid sequence of hnpfk (SEQ ID NO:3). 
     
     
         8 . The capture agent of  claim 5 , wherein the secondary ligand comprises the amino acid sequence of eihny (SEQ ID NO:4). 
     
     
         9 . The capture agent of  claim 1 , wherein the anchor ligand and secondary ligand are linked together via a 1,4-substituted-1,2,3-triazole residue (Tz4) or via a 1,5-substituted-1,2,3-triazole residue (Tz5). 
     
     
         10 . The capture agent of  claim 1 , wherein the capture agent is labeled with a label selected from the group consisting of biotin, copper-DOTA, biotin-PEG3, aminooxyacetate,  19 FB,  18 FB and FITC-PEG3. 
     
     
         11 . The capture agent of  claim 1 , wherein the capture agent is labeled with the detectable moiety selected from the group consisting of  6 Cu DOTA,  68 Ga DOTA,  68 Ga NOTA,  18 F, Al 18 F NOTA,  64 Cu,  68 Ga,  89 Zr,  124 I,  86 Y,  94m Tc,  110m In  11 C and  76 Br. 
     
     
         12 . The capture agent of  claim 1 , wherein the capture agent has the structure: 
       
         
           
           
               
               
           
         
       
     
     
         13 . The capture agent of  claim 1 , wherein the capture agent has the structure: 
       
         
           
           
               
               
           
         
       
     
     
         14 . The capture agent of  claim 1 , wherein the capture agent has having the structure: 
       
         
           
           
               
               
           
         
       
     
     
         15 . A composition comprising two or more capture agents of  claim 1 . 
     
     
         16 . A method for detecting anti-gp41 antibodies in a biological sample, comprising the step of administering to the biological sample one or more capture agents of  claim 1 . 
     
     
         17 . A method for detecting anti-gp41 antibodies in a biological sample, comprising the step of administering to the biological sample one or more capture agents of  claim 10 . 
     
     
         18 . A method for detecting anti-gp41 antibodies in a biological sample using an immunoassay, wherein the immunoassay utilizes a capture agent of  claim 1 . 
     
     
         19 . A method of diagnosing an HIV infection in a subject, the method comprising the steps of:
 a) administering to a biological sample from the subject one or more capture agents of  claim 1 , wherein each capture agent is linked to a detectable moiety;   b) binding antibody in the biological sample to a substrate; and   c) detecting the moiety linked to the capture agent on the substrate; wherein detection of the moiety on the substrate diagnoses an HIV infection in the subject.   
     
     
         20 . A method of monitoring treatment of HIV infection in a subject, comprising the steps of:
 a) administering to a first biological sample from the subject one or more capture agents of  claim 1 , wherein each capture agent is linked to a detectable moiety;   b) detecting the moiety linked to the capture agent bound to an anti-gp41 antibody;   c) administering a treatment for HIV to the subject;   d) administering to a second biological sample from the subject one or more capture agents of  claim 1 , wherein each capture agent is linked to a detectable moiety; and   e) detecting the moiety linked to the capture agent bound to an anti-gp41 antibody;   wherein if less of the moiety is detected in the second biological sample than in the first biological sample the treatment is improving the HIV infection in the subject.   
     
     
         21 . A method of detecting anti-gp41 antibodies in a biological sample, comprising the steps of:
 a) exposing the sample to a capture agent of  claim 1 , wherein the capture agent is linked to a detectable moiety; and   b) detecting the moiety linked to the capture agent bound to the anti-gp41 antibody in the sample, wherein detection of the moiety indicates the presence of anti-gp41 antibodies in the subject.   
     
     
         22 . A multiplex capture agent comprising two or more capture agents of  claim 1 , wherein the multiplex capture agent specifically binds two or more anti-gp41 antibodies.