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.

1 - 54 . (canceled) 
     
     
         55 . A modified liposome comprising: a) one or more lipids found in a host cell plasma membrane lipid bilayer in a region of the lipid raft or one or more types of lipids normally found in HIV-1; b) an adjuvant, c) a protein or peptide epitope from HIV-1 virus and d) nef or nef with env antigens, wherein said modified liposome produces monoclonal antibodies having specificity for both the lipid epitopes and the protein or peptide epitope. 
     
     
         56 . The modified liposome of  claim 55 , wherein said lipid epitopes comprise one or more of phosphatidylethanolamine, sphingomyelin, phosphatidylserine, phosphatidylinositol-4-phosphate, phosphatidylinositol, GalCer, SGalCer, CTH, GM1, and GM3. 
     
     
         57 . The modified liposome of  claim 55 , wherein said protein or peptide epitopes are from HIV-1 and comprise one or more of gp160, gp 140, gp120 and gp41. 
     
     
         58 . The modified liposome of  claim 55 , wherein the proteins or peptide epitope are attached to the surface of the liposomes, intercalated into a liposomal membrane bilayer, or encapsulated in an aqueous space inside of the liposome. 
     
     
         59 . The modified liposome of  claim 55 , wherein the adjuvant is Lipid A. 
     
     
         60 . A modified liposome comprising:
 a. lipid epitopes to HIV-1, wherein said lipid epitopes comprise one or more of phosphatidylethanolamine, sphingomyelin, phosphatidylserine, phosphatidylinositol-4-phosphate, phosphatidylinositol, GalCer, SGalCer, CTH, GM1 and GM3;   b. an adjuvant;   c. a protein or peptide epitopes from HIV-1 virus, wherein said protein or peptide epitopes are from HIV-1 comprise one or more of gp160, gp 140, gp120 and gp41; and   d. nef or nef with env antigens,   wherein said modified liposome has a property that it is capable of producing monoclonal antibodies having specificity for both the lipid and the HIV protein or peptide.   
     
     
         61 . The modified liposome of  claim 60 , wherein the adjuvant is Lipid A. 
     
     
         62 . A method of making liposomes that that have a property that they can produce monoclonal antibodies having specificity for both HIV lipid and HIV protein or peptide, said method comprising the steps of:
 a. obtaining liposomes having one or more lipids found in a host cell plasma membrane lipid bilayer in a region of a lipid raft or one or more types of lipids normally found in HIV-1;   b. modifying said liposomes by incorporating: (1) an adjuvant, (2) a protein or peptide epitope from HIV-1 virus, and (3) nef or nef with env antigens.   
     
     
         63 . The method of  claim 62 , wherein said lipids comprise one or more of phosphatidylethanolamine, sphingomyelin, phosphatidylserine, phosphatidylinositol-4-phosphate, phosphatidylinositol, GalCer, SGalCer, CTH, GM1 and GM3. 
     
     
         64 . The method of  claim 62 , wherein said protein or peptide epitopes are from HIV-1 and comprise one or more of gp160, gp 140, gp120 and gp41. 
     
     
         65 . The method of  claim 62 , wherein said adjuvant is Lipid A. 
     
     
         66 . A vaccine comprising the modified liposome of  claim 55 . 
     
     
         67 . A vaccine comprising the modified liposome of  claim 60 . 
     
     
         68 . An isolated dual- or multi-specific monoclonal antibody that has an antigen binding site that is dual- or multi-specific in binding both lipid and lipid associated HIV-1 protein antigen, wherein said monoclonal antibody was made by the following process:
 a) obtaining liposomes having lipid epitopes to HIV-1 and modifying said organized lipid structure by incorporating (1) an adjuvant and (2) a protein or peptide epitope from HIV-1 virus;   b) immunizing a mammal with said liposomes;   c) producing said antibodies, wherein said antibodies have simultaneous recognition subsites to said lipid epitopes in said liposome and to said protein or peptide epitope to HIV-1 virus;   d) identifying said antibodies with simultaneous recognition subsites to said lipid epitopes in said liposomes and to said protein or peptide epitope of said HIV-1 virus;   e) isolating only said antibodies with simultaneous recognition subsites to said lipid epitopes in said liposome and to said protein or peptide epitope of said HIV-1; 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 HIV-1 epitope.   
     
     
         69 . 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.   
     
     
         70 . The method of  claim 69 , wherein said lipid and protein or peptide epitopes are from the same entity. 
     
     
         71 . The method of  claim 70  wherein said same entity is selected from the group consisting of viruses, bacteria, hormones, fungi, cancer cells and protozoa. 
     
     
         72 . The method of  claim 69 , wherein said adjuvant is Lipid A. 
     
     
         73 . The method of  claim 69 , wherein said organized lipid structure is a lipid or liposome. 
     
     
         74 . The method of  claim 69 , 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. 
     
     
         75 . 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.   
     
     
         76 . The method of  claim 75 , 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. 
     
     
         77 . The method of  claim 75 , 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. 
     
     
         78 . 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. 
     
     
         79 . The method of  claim 78 , wherein the anionic phospholipids is phosphatidylinositol phosphate (PIP). 
     
     
         80 . The method of  claim 78 , wherein said anionic phospholipids is cardiolipin. 
     
     
         81 . 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.   
     
     
         82 . A method of binding PIP antigen and or cardiolipin (CL) comprising:
 administering anti-PIP antibody of  claim 81  to a medium containing PIP antigen or CL antigen.   
     
     
         83 . 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 81 .