Glycopeptide and uses thereof

Abstract

A glycolipopeptide comprising a carbohydrate component, a peptide component and a lipid component, for use as a therapeutic or prophylactic vaccine. Also provided are monoclonal and polyclonal antibodies that recognize the glycolipopeptide of the invention, as well as uses thereof.

1 - 51 . (canceled) 
     
     
         52 . A glycolipopeptide comprising:
 at least one carbohydrate component comprising a B-epitope;   at least one peptide component comprising a T-epitope; and   at least one lipid component;   
       wherein the carbohydrate component and the peptide component are heterologous with respect to each other. 
     
     
         53 . The glycolipopeptide of  claim 52  wherein the B-epitope is from a microorganism. 
     
     
         54 . The glycolipopeptide of  claim 53  wherein the microorganism is selected from the group consisting of a virus, a bacterium, a fungus, and a protozoan. 
     
     
         55 . The glycolipopeptide of  claim 53  wherein the microorganism is a human immunodeficiency virus or a hepatitis C virus. 
     
     
         56 . The glycolipopeptide of  claim 52  wherein the B epitope is overexpressed on a cancer cell. 
     
     
         57 . The glycolipopeptide of  claim 52  wherein the carbohydrate component comprises a self-antigen. 
     
     
         58 . The glycolipopeptide of  claim 57  wherein the self-antigen comprises a MUC-1 glycopeptide. 
     
     
         59 . The glycolipopeptide of  claim 52  wherein the carbohydrate component comprises a viral antigen. 
     
     
         60 . The glycolipopeptide of  claim 59  wherein the viral antigen is from human immunodeficiency virus or a hepatitis C virus. 
     
     
         61 . The glycolipopeptide of  claim 52  wherein the carbohydrate component comprises a glycoconjugate selected from the group consisting of a glycosylated protein, a glycosylated peptide, a glycosylated lipid, a glycosylated amino acid, a DNA and an RNA. 
     
     
         62 . The glycolipopeptide of  claim 61  wherein the glycosylated peptide or glycosylated protein comprises a β-N-acetylglucosamine (β-O-GlcNAc) modified peptide. 
     
     
         63 . The glycolipopeptide of  claim 52  comprising a glycosylated peptide, wherein the glycosylated peptide comprises the B-epitope and the T-epitope. 
     
     
         64 . The glycolipopeptide of  claim 52  wherein the carbohydrate component comprises a heparan sulfate fragment. 
     
     
         65 . The glycolipopeptide of  claim 52  wherein the lipid component comprises a Toll-like receptor (TLR) ligand. 
     
     
         66 . The glycolipopeptide of  claim 52  wherein the lipid component facilitates internalization of the glycolipopeptide by a target cell. 
     
     
         67 . The glycolipopeptide of  claim 52  wherein the T-epitope comprises a helper T-epitope. 
     
     
         68 . A pharmaceutical composition comprising:
 at least one of the glycolipopeptide of  claim 52  or a monoclonal or polyclonal antibody that binds thereto; and   a pharmaceutically acceptable carrier.   
     
     
         69 . The pharmaceutical composition of  claim 68  further comprising a liposome. 
     
     
         70 . The pharmaceutical composition of  claim 69  wherein the glycolipopeptide is covalently or noncovalently incorporated into the liposome. 
     
     
         71 . The pharmaceutical composition of  claim 70  wherein a plurality of glycolipopeptides are covalently or noncovalently incorporated in the liposome. 
     
     
         72 . The pharmaceutical composition of  claim 71  wherein at least two of the plurality of glycolipopeptides contain different B-epitopes. 
     
     
         73 . The pharmaceutical composition of  claim 71  wherein at least two of the plurality of glycolipopeptides contain the same B-epitopes. 
     
     
         74 . The pharmaceutical composition of  claim 68  which does not contain an external adjuvant. 
     
     
         75 . The pharmaceutical composition of  claim 68  further comprising an external adjuvant. 
     
     
         76 . The pharmaceutical composition of  claim 75  wherein the external adjuvant comprises QS-21. 
     
     
         77 . The pharmaceutical composition of  claim 68  formulated for use as a vaccine. 
     
     
         78 . A method for treating or preventing an infection, disease or disorder in a subject comprising administering to the subject the pharmaceutical composition of  claim 68 . 
     
     
         79 . The method of  claim 78  wherein the pharmaceutical composition includes an adjuvant comprising QS-21, and wherein the inclusion of QS-21 skews the immune response of the subject toward a Th1 response, compared to a comparable pharmaceutical composition that does not include QS-21. 
     
     
         80 . The method of  claim 78  wherein the infection, disease or disorder is caused by a microorganism. 
     
     
         81 . The method of  claim 80  wherein the microorganism is selected from the group consisting of a virus, a bacterium, a fungus, and a protozoan. 
     
     
         82 . The method of  claim 78  wherein the infection, disease or disorder is caused by a human immunodeficiency virus or a hepatitis C virus. 
     
     
         83 . The method of  claim 78  wherein the disease is cancer, a precancerous condition, or an autoimmune disease. 
     
     
         84 . A polyclonal or monoclonal antibody that binds to the glycolipopeptide of  claim 52 . 
     
     
         85 . The polyclonal or monoclonal antibody of  claim 84  which is an IgG antibody. 
     
     
         86 . The polyclonal or monoclonal antibody of  claim 85  which has broad selectivity for O-GlcNAc modified proteins. 
     
     
         87 . A hybridoma that produces a monoclonal antibody that binds to the glycolipopeptide of  claim 52 . 
     
     
         88 . A method for making a glycolipopeptide comprising:
 contacting a candidate compound with a target cell containing a Toll-like receptor (TLR);   determining whether the candidate compound binds to the TLR so as to identify a TLR ligand; and   covalently linking the identified TLR ligand to a carbohydrate component comprising a B-epitope and a peptide component comprising a T-epitope to yield a glycolipopeptide comprising the carbohydrate component, the peptide component, and the TLR ligand.   
     
     
         89 . The method of  claim 88  further comprising determining whether the candidate compound is internalized by the target cell. 
     
     
         90 . The method of  claim 88  wherein the candidate compound comprises a lipid. 
     
     
         91 . A glycolipopeptide made according to the method of  claim 88 . 
     
     
         92 . A kit comprising:
 a monoclonal antibody that binds to the glycolipopeptide of  claim 52 ;   packaging; and   instructions for use.   
     
     
         93 . The kit of  claim 92  wherein the monoclonal antibody is conjugated to a detectable label. 
     
     
         94 . The kit of  claim 92  further comprising a second antibody that binds to the monoclonal antibody. 
     
     
         95 . The kit of  claim 94  wherein the second antibody is conjugated to a detectable label. 
     
     
         96 . The kit of  claim 92  formulated for diagnostic or laboratory research. 
     
     
         97 . A method for detecting, diagnosing, or monitoring an infection, disease or disorder in a subject, the method comprising:
 contacting a biological sample from the subject with an antibody that binds to the glycolipopeptide of  claim 52 ; and   detecting binding of the antibody to a component in the biological sample;   
       wherein binding of the antibody to a sample component is indicative of the presence of the infection, disease or disorder in the subject. 
     
     
         98 . The method of  claim 97  further comprising:
 quantitating the level of antibody binding to the sample component; 
 quantitating the level of antibody binding to components in a comparable non-diseased sample; and 
 comparing the binding levels; 
 
       wherein a change in antibody binding in the biological sample compared to the non-diseased sample is indicative of the presence of the infection, disease or disorder in the subject. 
     
     
         99 . A method for detecting a glycosylated protein comprising:
 contacting a biological sample with an antibody that binds to a glycolipopeptide of  claim 52 ; and   detecting binding of the antibody to the protein.   
     
     
         100 . The method of  claim 99  further comprising identifying the protein. 
     
     
         101 . A method for identifying a protein associated with a disease state comprising:
 contacting a first biological sample associated with a disease state with an antibody that binds to a glycolipopeptide of  claim 52 ;   contacting a second biological sample associated with different disease state or no disease with said antibody;   detecting binding of said antibody to glycosylated proteins in the first and second samples; and   identifying glycosylated proteins that are enriched in one sample compared to the other;   
       wherein a difference in the amount of a glycosylated protein in the two samples is indicative of a protein associated with a disease state. 
     
     
         102 . The method of  claim 101  further comprising identifying the protein associated with a disease state. 
     
     
         103 . A glycolipopeptide comprising:
 at least one carbohydrate component comprising a B-epitope;   at least one peptide component comprising a T-epitope; and   at least one lipid component;   
       wherein the carbohydrate component comprises a saccharide selected from the group consisting of N-acetylglucosamine (GlcNAc), N-acetylgalactoseamine (GalNAc), mannose, or a glycosaminoglycan or fragment thereof. 
     
     
         104 . The glycolipopeptide of  claim 103  wherein the saccharide is O-linked, S-linked or N-linked to the glycolipopeptide. 
     
     
         105 . The glycolipopeptide of  claim 103  wherein the saccharide comprises β-N-acetylglucosamine (β-O-GlcNAc). 
     
     
         106 . The glycolipopeptide of  claim 103  wherein the glycosaminoglycan is selected from the group consisting of heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate and hyaluronan. 
     
     
         107 . The glycolipopeptide of  claim 103  wherein the carbohydrate component comprises a glycopeptide comprising the saccharide. 
     
     
         108 . The glycolipopeptide of  claim 103  wherein the carbohydrate component comprises a self-antigen. 
     
     
         109 . The glycolipopeptide of  claim 103  wherein the lipid component comprises a Toll-like receptor (TLR) ligand. 
     
     
         110 . A polyclonal or monoclonal antibody that binds to the glycolipopeptide of  claim 103 . 
     
     
         111 . The polyclonal or monoclonal antibody of  claim 110  which is an IgG antibody. 
     
     
         112 . The polyclonal or monoclonal antibody of  claim 111  which binds a broad spectrum of glycoproteins. 
     
     
         113 . A hybridoma that produces a monoclonal antibody that binds to the glycolipopeptide of  claim 103 . 
     
     
         114 . The polyclonal or monoclonal antibody of  claim 110  that binds to glycolipopeptide 52. 
     
     
         115 . A pharmaceutical composition comprising:
 the glycolipopeptide of  claim 103 , or a polyclonal or monoclonal antibody that binds thereto; and   a pharmaceutically acceptable carrier.   
     
     
         116 . The pharmaceutical composition of  claim 115  comprising a monoclonal antibody. 
     
     
         117 . A kit comprising:
 a monoclonal antibody that binds to the glycolipopeptide of  claim 103 ;   packaging; and   instructions for use.   
     
     
         118 . The kit of  claim 117  wherein the antibody is conjugated to a detectable label. 
     
     
         119 . The kit of  claim 117  further comprising a second antibody that binds to the first antibody. 
     
     
         120 . The kit of  claim 119  wherein the second antibody is conjugated to a detectable label. 
     
     
         121 . The kit of  claim 117  formulated for diagnostic or laboratory research use. 
     
     
         122 . A method for detecting, diagnosing or monitoring an infection, disease or disorder in a subject, the method comprising:
 contacting a biological sample from the subject with an antibody that binds to the glycolipopeptide of  claim 103 ; and   detecting binding of the antibody to a component in the biological sample;   
       wherein binding of the antibody to a sample component is indicative of the presence of the infection, disease or disorder in the subject. 
     
     
         123 . The method of  claim 122  further comprising:
 quantitating the level of antibody binding to the sample component; 
 quantitating the level of antibody binding to components in a comparable non-diseased sample; and 
 comparing the binding levels; 
 wherein a change in antibody binding in the biological sample compared to the non-diseased sample is indicative of the presence of the infection, disease or disorder in the subject. 
 
     
     
         124 . A method for detecting a glycosylated protein comprising:
 contacting a biological sample with an antibody that binds to a glycolipopeptide of  claim 103 ; and   detecting binding of the antibody to the protein.   
     
     
         125 . The method of  claim 124  further comprising identifying the protein. 
     
     
         126 . A method for identifying a protein associated with a disease state comprising:
 contacting a first biological sample associated with a disease state with an antibody that binds to a glycolipopeptide of  claim 103 ;   contacting a second biological sample associated with different disease state or no disease with said antibody;   detecting binding of said antibody to glycosylated proteins in the first and second samples; and   identifying glycosylated proteins that are enriched in one sample compared to the other;   
       wherein a difference in the amount of a glycosylated protein in the two samples is indicative of a protein associated with a disease state. 
     
     
         127 . A polyclonal or monoclonal pan-specific O-GlcNAc antibody. 
     
     
         128 . The antibody of  claim 127  which in an IgG antibody. 
     
     
         129 . The antibody of  claim 127  which binds to the glycolipopeptide of  claim 103 . 
     
     
         130 . The method of  claim 126  further comprising identifying the protein associated with a disease state. 
     
     
         131 . The method of  claim 126  wherein the antibody is a pan-specific O-GlcNAc antibody. 
     
     
         132 . The kit of  claim 117  wherein the antibody is a pan-specific O-GlcNAc antibody. 
     
     
         133 . A method for analyzing a biological sample comprising different glycoproteins, the method comprising:
 contacting the biological sample with a pan-specific O-GlcNAc antibody that binds to the glycolipopeptide  claim 103 ; and   detecting binding of the antibody to different glycoproteins in the biological sample.   
     
     
         134 . The method of  claim 133  further comprising determining the O-GlcNAc level in the biological sample. 
     
     
         135 . The method of  claim 134  wherein O-GlcNAc levels are determined before and after a treatment intervention, over time to monitor progression of disease, or to compare normal samples with samples from patients suspected of suffering from a disease, infection or disorder characterized by changes in protein glycosylation. 
     
     
         136 . The method of  claim 133  further comprising isolating, identifying and/or characterizing a glycoprotein bound by the pan-specific O-GlcNAc antibody. 
     
     
         137 . The method of  claim 136  wherein the glycoprotein has altered glycosylation in a disease state. 
     
     
         138 . The method of  claim 135  wherein the disease is Alzheimer's disease or Type II diabetes. 
     
     
         139 . The method of  claim 133  wherein the different glycoproteins recognized by the pan-specific antibodies share a substantially similar or identical glycopeptide sequence and/or a substantially similar secondary or tertiary structure at the glycosylation site. 
     
     
         140 . The method of  claim 133  wherein the pan-specific O-GlcNAc antibody has a broad selectivity for O-GlcNAc modified proteins and/or recognizes a broad spectrum of glycoproteins. 
     
     
         141 . The method of  claim 133  further comprising determining the site localization for an O-GlcNAc modification.