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US 2003/0211117 A1Published application
Synthetic antigen for the detection of aids-related disease
OwnerWesley Loren Cosandinventor
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Abstract
Novel peptides are provided having substantially the same sequence as immunologically significant fragments of AIDS-related viruses. The polypeptides can be used as reagents in the determination of exposure of a human host to the virus. Of particular interest is the use of polypeptides in screening blood products.
Claims36
In a method for detecting the presence of LAV/HTLV-III virus or antibody to LAV/HTLV-III virus where a sample is combined with a composition having epitopic sites immunologically competitive with LAV/HTLV-III epitopic sites, whereby antibodies bind to such protein composition to form a specific binding pair complex and the amount of complex formation is determined, the improvement which comprises:
- employing in the assay medium as a reagent a compostion, containing at least one peptide which has at least five amino acids and fewer than 50 amino acids and is encoded for by the coding region of LAV/HTLV-III from bp 900 to bp 1421 or bp 7210 to bp 7815.
In a method for detecting the presence of LAV/HTLV-III virus or antibody to LAV/HTLV-III virus where a sample is combined with a composition having epitopic sites immunologically competitive with LAV/HTLV-III epitopic sites, whereby antibodies bind to each such protein composition to form at least one specific binding pair complex and the amount of complex formation is determined, the improvement which comprises:
- employing in the assay medium as a reagent a composition, containing at least one peptide which has at least five amino acids in a sequence which comes within the sequence of at least one of the following peptide sequences:
- 24 (I) (15) Y-Asp-Cys-Lys-Thr-Ile-Leu-Lys-Ala-Leu- Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Met-Met- Thr-Ala-Cys-X (II) (17) Y-Leu-Lys-Glu-Thr-Ile-Asn-Glu-Glu-Ala- Ala-Glu-Trp-Asp-Arg-Val-His-Pro-Val-His- Ala-Z-X (III) (92) Y-Asp-Arg-Val-His-Pro-Val-His-Ala-Gly-Pro-Ile-Ala-Pro- Gly-Gln-X (IV) (90) Y-Tyr-Ser-Pro-Thr-Ser-Ile-Leu-Asp-Ile-Arg- Gln-Gly-Pro-Lys-Glu-Pro-Phe-Arg-Asp-Tyr-Val- Asp-Arg-Phe-Tyr-Lys-Thr-Leu-Arg-Z-X (V) (88) Y-Asn-Trp-Nor-Thr-Glu-Thr-Leu-Leu-Val-Gln- Asn-Ala-Asn-Pro-Asp-Cys-Lys-Thr-Ile-Leu-Lys- Ala-Leu-Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Nor- Nor-Thr-Ala-Cys-X (VI) (97) Y-Arg-Glu-Leu-Glu-Arg-Phe-Ala-Val-Asn-Pro-Gly- Leu-Leu-Glu-Thr-Ser-Glu-Gly-Cys-Arg-Gln-Ile- Leu-Gly-Gln-Leu-Gln-Pro-Ser-Leu-Gln-Thr-X (VII) (71) Y-Asp-Thr-Gly-His-Ser-Ser-Gln-Val-Ser-Gln- Asn-Tyr (VIII) (36) Val-Lys-Ile-Glu-Pro-Leu-Gly-Val-Ala-Pro- Thr-Lys-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg- Glu-Lys-Arg-Ala-Z-X (IX) (56) Ile-Lys-Gln-Leu-Gln-Ala-Arg-Ile-Leu- Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Z-X (X) (39) Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys- Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys- Ser-Gly-Lys-Leu-Ile-Cys-X (XI) (40) Y-Lys-Ser-Leu-Glu-Gln-Ile-Trp-Asn-Asn- Met-Thr-Trp-Met-Glu-Trp-Asp-Arg-Glu- Ile-Asn-Z-X (XII) (23) Y-His-Ser-Leu-Ile-Glu-Glu-Ser-Gln-Asn- Gln-Gln-Glu-Lys-Asn-Glu-Gln-Glu-Leu-Leu- Glu-Leu-Asp-Lys-Trp-Z-X (XIII) (79) Y-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly- Cys-Ser-Gly-Lys-Leu-Ile-Cys-X,
- where X is OH or NH2, and Y and Z, when present, are amino acids added to facilitate coupling, and where said peptide is free of other peptides or conjugated to a macromolecule for which antibodies in human sera are substantially absent.
A method for determining the presence of antibodies to LAV/HTLV-III in a physiological fluid, said method comprising:
- introducing a human serum, plasma or blood sample into a sample container coated at least in part with at least one peptide having at least five amino acids which comes within the sequence of at least one of the following peptide sequences:
- 26 (I) (15) Y-Asp-Cys-Lys-Thr-Ile-Leu-Lys-Ala-Leu- Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Met-Met- Thr-Ala-Cys-X (II) (17) Y-Leu-Lys-Glu-Thr-Ile-Asn-Glu-Glu-Ala- Ala-Glu-Trp-Asp-Arg-Val-His-Pro-Val-His- Ala-X (III) (92) Y-Asp-Arg-Val-His-Pro-Val-His-Ala-Gly-Pro Ile-Ala-Pro-Gly-Gln-X (IV) (90) Y-Tyr-Ser-Pro-Thr-Ser-Ile-Leu-Asp-Ile-Arg Gln-Gly-Pro-Lys-Glu-Pro-Phe-Arg-Asp-Tyr-Val- Asp-Arg-Phe-Tyr-Lys-Thr-Leu-Arg-Z-X (V) (88) y-Asn-Trp-Nor-Thr-Glu-Thr-Leu-Leu-Val-Gln- Asn-Ala-Asn-Pro-Asp-Cys-Lys-Thr-Ile-Leu-Lys- Ala-Leu-Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Nor- Nor-Thr-Ala-Cys-X (VI) (97) Y-Arg-Glu-Leu-Glu-Arg-Phe-Ala-Val-Asn-Pro-Gly- Leu-Leu-Glu-Thr-Ser-Glu-Gly-Cys-Arg-Gln-Ile- Leu-Gly-Gln-Leu-Gln-Pro-Ser-Leu-Gln-Thr-X (VII) (71) Y-Asp-Thr-Gly-His-Ser-Ser-Gln-Val-Ser-Gln- Asn-Tyr (VIII) (36) Val-Lys-Ile-Glu-Pro-Leu-Gly-Val-Ala-Pro Thr-Lys-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg Glu-Lys-Arg-Ala-Z-X (IX) (56) Ile-Lys-Gln-Leu-Gln-Ala-Arg-Ile-Leu- Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Z-X (X) (39) Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys- Ser-Gly-Lys-Leu-Ile-Cys-X (XI) (40) Y-Lys-Ser-Leu-Glu-Gln-Ile-Trp-Asn-Asn- Met-Thr-Trp-Met-Glu-Trp-Asp-Arg-Glu- Ile-Asn-Z-X (XII) (23) Y-His-Ser-Leu-Ile-Glu-Glu-Ser-Gln-Asn- Gln-Gln-Glu-Lys-Asn-Glu-Gln-Glu-Leu-Leu- Glu-Leu-Asp-Lys-Trp-Z-X (XIII) (79) Y-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly- Cys-Ser-Gly-Lys-Leu-Ile-Cys-X,
- where X is OH or NH2, and Y and Z, if present, are amino acids added to facilitate coupling, wherein said peptides are free of other peptides or conjugated to a macromolecule for which antibodies in human sera are substantially absent;
- incubating for a sufficient time for complex formation to occur;
- and
- determining the formation of complex by employing a labeled specific binding protein which binds to said complex and provides a detectable signal.
3 more elements
A method for determining the presence of antibodies to LAV/HTLV-III in a physiological fluid, said method comprising:
- combining a human serum, plasma or blood sample with at least one labeled peptide having at least five amino acids in a sequence which comes within the sequence of at least one of the following peptide sequences:
- 27 (I) (15) Y-Asp-Cys-Lys-Thr-Ile-Leu-Lys-Ala-Leu- Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Met-Met Thr-Ala-Cys-X (II) (17) Y-Leu-Lys-Glu-Thr-Ile-Asn-Glu-Glu-Ala- Ala-Glu-Trp-Asp-Arg-Val-His-Pro-Val-His- Ala-X (III) (92) Y-Asp-Arg-Val-His-Pro-Val-His-Ala-Gly-Pro- Ile-Ala-Pro-Gly-Gln-X (IV) (90) Y-Tyr-Ser-Pro-Thr-Ser-Ile-Leu-Asp-Ile-Arg- Gln-Gly-Pro-Lys-Glu-Pro-Phe-Arg-Asp-Tyr-Val- Asp-Arg-Phe-Tyr-Lys-Thr-Leu-Arg-z-x (V) (88) Y-Asn-Trp-Nor-Thr-Glu-Thr-Leu-Leu-Val-Gln- Asn-Ala-Asn-Pro-Asp-Cys-Lys-Thr-Ile-Leu-Lys- Ala-Leu-Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Nor- Nor-Thr-Ala-Cys-X (VI) (97) Y-Arg-Glu-Leu-Glu-Arg-Phe-Ala-Val-Asn-Pro-Gly- Leu-Leu-Glu-Thr-Ser-Glu-Gly-Cys-Arg-Gln-Ile- Leu-Gly-Gln-Leu-Gln-Pro-Ser-Leu-Gln-Thr-X (VII) (71) Y-Asp-Thr-Gly-His-Ser-Ser-Gln-Val-Ser-Gln- Asn-Tyr (VIII) (36) Val-Lys-Ile-Glu-Pro-Leu-Gly-Val-Ala-Pro- Thr-Lys-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg Gln-Lys-Arg-Ala-Z-X (IX) (56) Ile-Lys-Gln-Leu-Gln-Ala-Arg-Ile-Leu- Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Z-X (X) (39) Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys- Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys- Ser-Gly-Lys-Leu-Ile-Cys-X (XI) (40) Y-Lys-Ser-Leu-Glu-Gln-Ile-Trp-Asn-Asn- Met-Thr-Trp-Met-Glu-Trp-Asp-Arg-Glu- Ile-Asn-Z-X (XII) (23) Y-His-Ser-Leu-Ile-Glu-Glu-Ser-Gln-Asn- Gln-Gln-Glu-Lys-Asn-Glu-Gln-Glu-Leu-Leu- Glu-Leu-Asp-Lys-Trp-Z-x (XIII) (79) Y-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly- Cys-Ser-Gly-Lys-Leu-Ile-Cys-X,
- where X is OH or NH2, and Y and Z, if present, are amino acids added to facilitate coupling, and said peptides are conjugated to a protein for which antibodies in human sera are substantially absent or unconjugated to a protein;
- incubating for a sufficient time for complex formation to occur;
- and
- determining the formation of complex as a result of change in a detectable signal resulting from complex formation.
3 more elements
A peptide of the formula:
- 28 (I) (15) Y-Asp-Cys-Lys-Thr-Ile-Leu-Lys-Ala-Leu- Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Met-Met- Thr-Ala-Cys-X,
- where X is OH or NH2, and Y, if present, is an amino acid added to facilitate coupling, N-terminal acetylated I, and I linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 29 (II) (17) Y-Leu-Lys-Glu-Thr-Ile-Asn-Glu-Glu-Ala Ala-Glu-Trp-Asp-Arg-Val-His-Pro-Val-His Ala-X,
- where X is OH or NH2, and Y, if present, is an amino acid added to facilitate coupling, N-terminal acetylated II, and II linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 30 (III) (92) Y-Asp-Arg-Val-His-Pro-Val-His-Ala-Gly-Pro- Ile-Ala-Pro-Gly-Gln-X,
- where X is OH or NH2, and Y, if present, is an amino acid added to facilitate coupling, N-terminal acetylated III, and III linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 31 (IV) (90) Y-Tyr-Ser-Pro-Thr-Ser-Ile-Leu-Asp-Ile-Arg Gln-Gly-Pro-Lys-Glu-Pro-Phe-Arg-Asp-Tyr-Val- Asp-Arg-Phe-Tyr-Lys-Thr-Leu-Arg-Z-X,
- where X is OH or NH2, and Y, if present, is an amino acid added to facilitate coupling, N-terminal acetylated IV, and IV linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 32 (V) (88) Y-Asn-Trp-Nor-Thr-Glu-Thr-Leu-Leu-Val-Gln- Asn-Ala-Asn-Pro-Asp-Cys-Lys-Thr-Ile-Leu-Lys- Ala-Leu-Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Nor- Nor-Thr-Ala-CyS-X,
- where X is OH or NH2, and Y, if present, is an amino acid added to facilitate coupling, N-terminal acetylated V, and V linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 33 (VI) (97) Y-Arq-Glu-Leu-Glu-Arg-Phe-Ala-Val-Asn-Pro-Gly- Leu-Leu-Glu-Thr-Ser-Glu-Gly-Cys-Arg-Gln-Ile- Leu-Gly-Gln-Leu-Gln-Pro-Ser-Leu-Gln-Thr-X,
- where X is OH or NH2, and Y, if present, is an amino acid added to facilitate coupling, N-terminal acetylated VI, and VI linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 34 (VII) (71) Y-Asp-Thr-Gly-His-Ser-Ser-Gln-Val-Ser-Gln- Asn-Tyr
- where Y, if present, is an amino acid added to facilitate coupling, N-terminal acetylated VII, and VII linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 35 (VIII) (36) Val-Lys-Ile-Glu-Pro-Leu-Gly-Val-Ala-Pro- Thr-Lys-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg- Glu-Lys-Arg-Ala-Z-X,
- where X is OH or Nil2, and Z, if present, is an amino acid added to facilitate coupling, N-terminal acetylated VIII, and VIII linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 36 (IX) (56) Ile-Lys-Gln-Leu-Gln-Ala-Arg-Ile-Leu- Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Z-X,
- where X is OH or NH2, and Z, if present, is an amino acid added to facilitate coupling, N-terminal acetylated IX, and IX linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 37 (X) (39) Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys- Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys- Ser-Gly-Lys-Leu-Ile-Cys-X,
- where X is OH or NH2, N-terminal acetylated X, and X linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 38 (XI) (40) Y-Lys-Ser-Leu-Glu-Gln-Ile-Trp-Asn-Asn- Met-Thr-Trp-Met-Glu-Trp-Asp-Arg-Glu- Ile-Asn-Z-X,
- where X is OH or NH2, and each of Y and Z, if present, is an amino acid added to facilitate coupling, N-terminal acetylated XI, and XI linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 39 (XII) (23) Y-His-Ser-Leu-Ile-Glu-Glu-Ser-Gln-Asn- Gln-Gln-Glu-Lys-Asn-Glu-Gln-Glu-Leu-Leu- Glu-Leu-Asp-Lys-Trp-Z-X,
- where X is OH or NH2, and each of Y and Z, if present, is an amino acid added to facilitate coupling, N-terminal acetylated XII, and XII linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A peptide of the formula:
- 40 (XIII) (79) Y-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly- Cys-Ser-Gly-Lys-Leu-Ile-Cys-X,
- where X is OH or NH2, and each of Y and Z. If present, is an amino acid added to facilitate coupling, N-terminal acetylated XIII, and XIII linked to a peptide or protein of at least 5,000 molecular weight, which peptide or protein does not normally bind to antibodies present in a human host.
A vaccine composition comprising at least one peptide of the peptides:
- 41 (I) (15) Y-Asp-Cys-Lys-Thr-Ile-Leu-Lys-Ala-Leu- Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Met-Met Thr-Ala-Cys-X (II) (17) Y-Leu-Lys-Glu-Thr-Ile-Asn-Glu-Glu-Ala- Ala-Glu-Trp-Asp-Arg-Val-His-Pro-Val-His- Ala-Z-X (III) (92) Y-Asp-Arg-Val-His-Pro-Val-His-Ala-Gly-Pro- Ile-Ala-Pro-Gly-Gln-X (IV) (90) Y-Tyr-Ser-Pro-Thr-Ser-Ile-Leu-Asp-Ile-Arg- Gln-Gly-Pro-Lys-Glu-Pro-Phe-Arg-Asp-Tyr-Val Asp-Arg-Phe-Tyr-Lys-Thr-Leu-Arg-Z-x (V) (88) Y-Asn-Trp-Nor-Thr-Glu-Thr-Leu-Leu-Val-Gln- Asn-Ala-Asn-Pro-Asp-Cys-Lys-Thr-Ile-Leu-Lys- Ala-Leu-Gly-Pro-Ala-Ala-Thr-Leu-Glu-Glu-Nor- Nor-Thr-Ala-Cys-X (VI) (97) Arg-Glu-Leu-Glu-Arg-Phe-Ala-Val-Asn-Pro-Gly- Leu-Leu-Glu-Thr-Ser-Glu-Gly-Cys-Arg-Gln-Ile- Leu-Gly-Gln-Leu-Gln-Pro-Ser-Leu-Gln-Thr (VII) (71) Asp-Thr-Gly-His-Ser-Ser-Gln-Val-Ser-Gln-Asn-Tyr (VIII) (36) Val-Lys-Ile-Glu-Pro-Leu-Gly-Val-Ala-Pro- Thr-Lys-Ala-Lys-Arg-Arg-Val-Val-Gln-Arg- Glu-Lys-Arg-Ala-Z-X (IX) (56) Ile-Lys-Gln-Leu-Gln-Ala-Arg-Ile-Leu- Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Z-X (X) (39) Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys- Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys- Ser-Gly-Lys-Leu-Ile-Cys-X (XI) (40) Y-Lys-Ser-Leu-Glu-Gln-Ile-Trp-Asn-Asn- Met-Thr-Trp-Met-Glu-Trp-Asp-Arg-Glu- Ile-Asn-Z-X (XII) (23) Y-His-Ser-Leu-Ile-Glu-Glu-Ser-Gln-Asn- Gln-Gln-Glu-Lys-Asn-Glu-Gln-Glu-Leu-Leu- Glu-Leu-Asp-Lys-Trp-Z-X (XIII) (79) Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly- Cys-Ser-Gly-Lys-Leu-Ile-Cys,
- where X is OH or NH2, and each of Y and Z. when present, are amino acids added to facilitate coupling,
- or said peptides conjugated to an immunogenic protein, said peptides or conjugates being present in an amount to provide an immunogenic response in a physiologically acceptable carrier.
A vaccine according to claim 34, wherein said peptides are coupled to immunogenic proteins.
Dependencies are inferred from the claim text.