Antibodies with simultaneous subsite specificities to protein and lipid epitopes

Abstract

Antibodies and method of making antibodies, either monoclonal or polyclonal wherein said antibodies have dual or multi-specific binding capacity to more than one type of antigenic epitope. The antibodies have simultaneous or independent recognition subsites to each of the epitopes. Antigenic epitopes include lipids, peptides, proteins, amino acid sequences, sugars and carbohydrates. Monoclonal antibodies and a method of making monoclonal antibodies of the invention include monoclonal antibodies that are broadly neutralizing to HIV-1 or other envelop viruses wherein the monoclonal antibody has subsites that simultaneously recognize protein and lipid epitopes from the virus.

What is claimed is: 
     
         1 . A method of making monoclonal antibodies that have an antigen binding site that is dual- or multi-specific in binding more than one type of antigenic epitope comprising:
 a) obtaining an organized lipid structure having a first lipid epitope and modifying said organized lipid structure by incorporating (1) an adjuvant and (2) a protein or peptide epitope;   b) immunizing a mammal with said organized lipid structure;   c) producing said antibodies, wherein said antibodies have simultaneous recognition subsites to said lipid epitopes in said organized lipid structure and to said protein or peptide epitope;   d) identifying said antibodies with simultaneous recognition subsites to said lipid epitopes in said organized lipid structure and to said protein or peptide epitope;   e) isolating only said antibodies with simultaneous recognition subsites to said lipid epitopes in said organized lipid structure and to said protein or peptide epitope; and   f) cloning said antibodies to obtain monoclonal dual-multi specific antibodies having dual or multi-specific antigen binding sites binding more than one antigenic epitope selected from the lipid epitope and the protein or peptide epitope.   
     
     
         2 . The method of  claim 1 , wherein said lipid and protein or peptide epitopes are from the same entity. 
     
     
         3 . The method of  claim 2  wherein said same entity is selected from the group consisting of viruses, bacteria, hormones, fungi, cancer cells and protozoa. 
     
     
         4 . The method of  claim 1 , wherein said adjuvant is Lipid A. 
     
     
         5 . The method of  claim 1 , wherein said organized lipid structure is a lipid or liposome. 
     
     
         6 . The method of  claim 1 , wherein said lipid epitope comprise one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, phosphatidylserine, phosphatidylinositol-4-phosphate, phosphatidylinositol, phosphatidyl glycerol, GalCer, SGalCer, CTH, GM1, GM3 and cholesterol. 
     
     
         7 . A method of making antibodies that have an antigen binding site that is dual- or multi-specific in binding more than one type of antigenic epitope comprising:
 a) obtaining liposomes having a first antigenic epitope;   b) modifying said liposomes by including a second antigenic epitope in said organized lipid structure;   c) immunizing a mammal with said liposomes;   d) producing said antibodies, wherein said antibodies have simultaneous recognition subsites to said first antigenic epitopes in said liposome and to said second antigenic epitopes;   e) identifying said antibodies with simultaneous recognition subsites to said first antigenic epitopes in said liposome and to said second antigenic epitope;   f) isolating only said antibodies with simultaneous recognition subsites to said first epitopes in said liposome and to said second antigenic epitope;   g). cloning said antibodies to obtain monoclonal dual-multi specific antibodies having dual or multi-specific antigen binding sites binding more than one antigenic epitope selected from said first and second antigenic epitopes.   
     
     
         8 . The method of  claim 7 , wherein said first antigenic epitope and second antigenic epitope is selected from the group consisting of protein, peptide, polypeptide, amino acid sequence, lipid, sugar and carbohydrate. 
     
     
         9 . The method of  claim 7 , wherein said lipid epitope comprise one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, phosphatidylserine, phosphatidylinositol-4-phosphate, phosphatidylinositol, phosphatidyl glycerol, GalCer, SGalCer, CTH, GM1, GM3 and cholesterol. 
     
     
         10 . A monoclonal antibody comprising;
 an antibody having subsites that simultaneously recognize one or more epitopes selected from the group consisting of lipids, proteins, peptides, sugars, and carbohydrates.   
     
     
         11 . A monoclonal antibody made by the process of  claim 1 . 
     
     
         12 . The monoclonal antibody of  claim 10 , wherein said protein or peptide epitopes are from HIV-1 and comprise one or more of gp160, gp 140, gp120 and gp41. 
     
     
         13 . The monoclonal antibody of  claim 10 , wherein said lipid epitopes comprise one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, phosphatidylserine, phosphatidylinositol-4-phosphate, phosphatidylinositol, Phosphatidyl glycerol, GalCer, SGalCer, CTH, GM1, GM3 and cholesterol. 
     
     
         14 . The monoclonal antibody of  claim 10 , wherein said lipid epitopes comprise one or more of the lipid epitopes found in a lipid raft region of a plasma membrane of a host cell. 
     
     
         15 . A method of making a monoclonal antibody to anionic phospholipids comprising: incorporating said anionic phospholipids and lipid A into a liposome; inserting said liposome into a mammal wherein said mammal produces monoclonal antibodies to said phospholipids. 
     
     
         16 . The method of  claim 15 , wherein the anionic phospholipids is phosphatidylinositol phosphate (PIP). 
     
     
         17 . The method of  claim 15 , wherein said anionic phospholipids is cardiolipin. 
     
     
         18 . A method of making a monoclonal antibody to phosphatidylinositol phosphate (PIP) comprising:
 incorporating PIP and Lipid A into a liposome;   inserting said liposome into a mammal, wherein said mammal produces monoclonal antibodies to said PIP.   
     
     
         19 . A method of binding PIP antigen and or cardiolipin (CL) comprising:
 administering anti-PIP antibody of  claim 18  to a medium containing PIP antigen or CL antigen.   
     
     
         20 . A method of inhibition of infection of HIV-1 in primary cultures of peripheral blood mononuclear cells by HIV-1 comprising administering the anti PIP antibody of  claim 18 .