GLYCOPEPTIDE AND USES THEREOF
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.