US2006063149A1PendingUtilityA1

Compositions and methods for detecting pathogen infection

Individually held — no corporate assignee on recordPriority: Dec 23, 2003Filed: Sep 6, 2005Published: Mar 23, 2006

Est. expiryDec 23, 2023(expired)· nominal 20-yr term from priority

Inventors:Francois-Xavier Berthet Francesc Vayreda Casadevall Maria Cruz Sanz Maria Teresa Llop Garcia Angels Mor Olle

A61P 37/08A61P 37/04A61P 43/00A61P 31/00A61P 31/22A61P 29/00A61P 33/06A61P 31/14A61P 31/06A61P 31/20A61P 35/02A61P 31/04A61P 25/28A61P 35/00A61P 31/18A61P 27/16A61P 31/16G01N 2800/2821A61P 15/02G01N 2800/347A61P 11/16A61P 11/00G01N 2800/065A61K 38/164G01N 2333/924G01N 2469/20G01N 33/573C07K 14/20G01N 33/571G01N 33/56911G01N 33/56983A61P 1/04G01N 2500/02C12N 9/2462G01N 2333/20G01N 2800/24A61P 15/18A61P 13/12A61P 15/16A61P 1/02Y02A50/30

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References

Claims

Abstract

The present invention generally features therapeutic and diagnostic compositions and methods for increasing or decreasing the binding of a lysozyme polypeptide to a Treponema pallidum P17 polypeptide (Tp17) or a Tp17-like polypeptide. More particularly, the invention relates to compositions and methods for detecting, treating, or preventing a pathogen infection or a chronic disorder; and to binding assays using a Tp17-like polypeptide and a lysozyme polypeptide.

Claims

exact text as granted — not AI-modified

1 . A fragment of a Tp17-like polypeptide, comprising: a lysozyme binding motif comprising at least four amino acids, two of the four amino acids selected from the group consisting of Cys, Pro, His, and Arg, wherein the lysozyme binding motif is not CKPHDC (SEQ ID NO. 24).

2 . The fragment of claim 1 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Xaa1 Xaa2 Arg Xaa3 Xaa4 Xaa5 Cys (SEQ ID NO. 314), wherein Xaa1, Xaa2, Xaa3, Xaa4, or Xaa5 is any amino acid, is absent, or is a peptide bond.

3 . The fragment of claim 1 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Xaa1 Xaa2 Arg Xaa3 Xaa4 Xaa5 Cys (SEQ ID NO. 314), wherein Xaa1 is Pro, Ala, Val, or Ser; Xaa2 is Gln, Glu, or His; Xaa3 is Leu or Met; Xaa4 is Ser, Ala, or Gly; and Xaa5 is Ser, Val, Ala, Lys, or Cys.

4 . The fragment of claim 1 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Pro Xaa1 Arg Xaa2 Xaa3 Xaa4 Cys (SEQ ID NO. 315), wherein Xaa1 is Gln, Glu, or His; Xaa2 is Leu or Met; Xaa3 is Ser or Ala; Xaa4 is Ser, Ala, or Val.

5 . The fragment of claim 4 , wherein the lysozyme binding motif comprises an amino acid sequence selected from the group consisting of CPHRLSVC (SEQ ID NO. 316), CPHRLSSC (SEQ ID NO. 317), CPERLASC (SEQ ID NO. 318), CPERMASC (SEQ ID NO. 319), CPERLSSC (SEQ ID NO. 320), and CPQRLSSC (SEQ ID NO. 321).

6 . The fragment of claim 5 , wherein the lysozyme binding motif comprises amino acid sequence CPHRLSVC (SEQ ID NO. 316).

7 . The fragment of claim 1 , wherein the polypeptide is derived from the E2 envelope glycoprotein of a Hepatitis C virus.

8 . The fragment of claim 1 , wherein the lysozyme binding motif is affixed to a solid support.

9 . The fragment of claim 1 , wherein the polypeptide is linked to a detectable label.

10 . A substantially pure nucleic acid molecule encoding the fragment of claim 2 .

11 . A vector comprising the nucleic acid molecule of claim 10 .

12 . A host cell comprising the vector of claim 11 .

13 . A fusion protein comprising the fragment of claim 1 .

14 . A composition comprising a first polypeptide comprising at least a lysozyme binding motif and a second polypeptide, wherein the lysozyme binding motif comprises at least four amino acids, two of which are selected from the group consisting of Cys, Pro, His, and Arg, wherein the first polypeptide is not the Ivy polypeptide and the second polypeptide comprisies a lysozyme.

15 . The composition of claim 14 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Xaa1 Xaa2 Arg Xaa3 Xaa4 Xaa5 Cys (SEQ ID NO. 314), wherein Xaa1, Xaa2, Xaa3, Xaa4, or Xaa5 is any amino acid, is absent, or is a peptide bond.

16 . The composition of claim 14 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Xaa1 Xaa2 Arg Xaa3 Xaa4 Xaa5 Cys (SEQ ID NO. 314), wherein Xaa1 is Pro, Ala, Val, or Ser; Xaa2 is Gln, Glu, or His; Xaa3 is Leu or Met; Xaa4 is Ser, Ala, or Gly; and Xaa5 is Ser, Val, Ala, Lys, or Cys.

17 . The composition of claim 14 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Pro Xaa1 Arg Xaa2 Xaa3 Xaa4 Cys (SEQ ID NO. 315), wherein Xaa1 is Gln, Glu, or His; Xaa2 is Leu or Met; Xaa3 is Ser or Ala; Xaa4 is Ser, Ala, or Val.

18 . The composition of claim 14 , wherein the lysozyme binding motif comprises an amino acid sequence selected from the group consisting of CPHRLSVC (SEQ ID NO. 316), CPHRLSSC (SEQ ID NO. 317), CPERLASC (SEQ ID NO. 318), CPERMASC (SEQ ID NO. 319), CPERLSSC (SEQ ID NO. 320), and CPQRLSSC (SEQ ID NO. 321).

19 . The composition of claim 14 , wherein the lysozyme binding motif comprises an amino acid sequence of CPHRLSVC.

20 . The composition of claim 14 , wherein the first polypeptide is derived from the E2 envelope glycoprotein of a Hepatitis C virus.

21 . The composition of claim 14 , wherein the second polypeptide comprises the following amino acid sequence:

Xaa Xaa Xaa Xaa Xaa Xaa Cys Xaa
(SEQ ID NO: 28)

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Trp Xaa Cys Xaa

Xaa Xaa Xaa Glu Ser Xaa Xaa Xaa

Thr Xaa Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Ser Xaa Asp Tyr Gly

Xaa Xaa Gln Ile Asn Xaa Xaa Xaa

Trp Cys Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Xaa Cys Xaa Xaa

Xaa Cys Xaa Xaa Leu Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Xaa Cys Ala Lys

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Trp Xaa Xaa Trp Xaa

Xaa Xaa Cys Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Cys Xaa Xaa Xaa,

wherein Xaa is any amino acid or is absent.

22 . A method for detecting an immune response against a pathogen in a subject, the method comprising the steps of:
(a) contacting a biological sample from the subject with an exogenous lysozyme and a polypeptide comprising a lysozyme binding motif; and (b) detecting antibody binding to the polypeptide or to a polypeptide-lysozyme complex.

23 . The method of claim 22 , wherein the diagnostic assay is an agglutination assay.

24 . The method of claim 22 , wherein the lysozyme is contacted with the polypeptide comprising the lysozyme binding motif prior to, during, or after contacting the biological sample.

25 . The method of claim 22 , wherein the lysozyme is derived from human.

26 . The method of claim 22 , wherein the pathogen is Hepatitis C virus.

27 . A kit for detecting an immune response to a pathogen comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprises at least a lysozyme binding motif and the second polypeptide is a lysozyme.

28 . The kit of claim 27 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Xaa1 Xaa2 Arg Xaa3 Xaa4 Xaa5 Cys (SEQ ID NO. 314), wherein Xaa1, Xaa2, Xaa3, Xaa4, or Xaa5 is any amino acid, is absent, or is a peptide bond.

29 . The kit of claim 27 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Xaa1 Xaa2 Arg Xaa3 Xaa4 Xaa5 Cys (SEQ ID NO. 314), wherein Xaa1 is Pro, Ala, Val, or Ser; Xaa2 is Gln, Glu, or His; Xaa3 is Leu or Met; Xaa4 is Ser, Ala, or Gly; and Xaa5 is Ser, Val, Ala, Lys, or Cys.

30 . The kit of claim 27 , wherein the lysozyme binding motif comprises an amino acid sequence of Cys Pro Xaa1 Arg Xaa2 Xaa3 Xaa4 Cys (SEQ ID NO. 315), wherein Xaa1 is Gln, Glu, or His; Xaa2 is Leu or Met; Xaa3 is Ser or Ala; Xaa4 is Ser, Ala, or Val.

31 . The kit of claim 27 , wherein the lysozyme binding motif comprises an amino acid sequence selected from the group consisting of CPHRLSVC (SEQ ID NO. 316), CPHRLSSC (SEQ ID NO. 317), CPERLASC (SEQ ID NO. 318), CPERMASC (SEQ ID NO. 319), CPERLSSC (SEQ ID NO. 320), and CPQRLSSC (SEQ ID NO. 321).

32 . The composition of claim 27 , wherein the lysozyme binding motif comprises an amino acid sequence of CPHRLSVC (SEQ ID NO. 316).

33 . The kit of claim 27 , wherein the second polypeptide comprises the following amino acid sequence:

Xaa Xaa Xaa Xaa Xaa Xaa Cys Xaa
(SEQ ID NO: 28)

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Trp Xaa Cys Xaa

Xaa Xaa Xaa Glu Ser Xaa Xaa Xaa

Thr Xaa Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Ser Xaa Asp Tyr Gly

Xaa Xaa Gln Ile Asn Xaa Xaa Xaa

Trp Cys Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Xaa Cys Xaa Xaa

Xaa Cys Xaa Xaa Leu Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Xaa Cys Ala Lys

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Trp Xaa Xaa Trp Xaa

Xaa Xaa Cys Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Cys Xaa Xaa Xaa,

wherein Xaa is any amino acid or is absent.

34 . The kit of claim 27 , wherein the first or the second polypeptide is attached to a solid support.

35 . The kit of claim 34 , wherein the solid support is selected from the group consisting of a resin, a gel, a bead, a well, a column, a chip, a membrane, a matrix, a plate, and a filter device.

36 . The kit of claim 27 , wherein the first or the second polypeptide is linked to a detectable label.

37 . The kit of claim 27 , wherein the pathogen is Hepatitis C virus.

38 - 42 . (canceled)

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