Binding moieties for human parvovirus B19

• US 6,291,197 B1
• Filed: 09/26/2000
• Issued: 09/18/2001
• Est. Priority Date: 11/05/1998
• Status: Expired due to Term

• Current Assignee: Dyax Corporation (Shire plc)
• Original Assignee: Dyax Corporation (Shire plc)
• Inventors: Whelihan, E. Fayelle
• Primary Examiner(s): SCHEINER, LAURIE A
• Application Number:
  US09/669,271
• Time in Patent Office: 357 Days
• Field of Search: 435/7.1, 435/5, 435/7.92, 435/200.1, 435/199.1, 435/252.1, 435/320.1, 424/147.1, 536/23.1
• US Class Current: 435/7.92
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: G01N 2333/015 Parvoviridae, e.g. feline panleukopenia virus, human Parvovirus
• CPC Class Codes: G01N 33/56983 Viruses

Abstract

Methods for detecting human parvovirus B19 in and removing it from biological samples such as blood are disclosed, together with reagents suitable for the purpose comprising binding moieties that recognize human parvovirus B19 and/or B19-like polypeptide and form a binding complex therewith. Preferred polypeptide binding moieties are particularly disclosed.

Claims

1. A recombinant bacteriophage expressing exogenous DNA encoding a parvovirus binding peptide having an amino acid sequence selected from the group consisting of:

I. X₁-X₂-Cys-X₃-X₄-X₅-X₆-X₇-Cys-X₈-X₉ (SEQ ID NO;

     1),

wherein X₁ is Phe or Leu or is not present;

X₂ is Phe or Ser;

X₃ is Arg, Gln, Ser, His, Ala, Leu, or Gly;

X₄ is Phe, Tyr, Leu, or Trp;

X₅ is Trp or Phe;

X₆ is Tyr, Pro, or His;

X₇ is Gly, Asn, Ser, Phe, or Asp;

X₈ is His, Asp, Ser or Pro;

X₉ is Pro, Ala, Phe, His, or Asp or is not present,

II. X₁₀-Phe-Cys-X₁₁-X₁₂-Trp-X₁₃-X₁₄-X₁₅-Cys-X₁₆-X₁₇ (SEQ ID NO;

     2),

wherein X₁₀ is His, Ala, or Phe;

X₁₁ is His, Trp, or Ser;

X₁₂ is Phe or Leu;

X₁₃ is Phe, Pro, or His;

X₁₄ is Gly or His;

X₁₅ is Gly or Asn;

X₁₆ Pro, Leu, or Asp;

X₁₇ is His or Asp,

III. X₁₈-Cys-X₁₉-X₂₀-X₂₁-X₂₂-X₂₃-X₂₄-X₂₅-Cys-X₂₆ (SEQ ID NO;

     3),

wherein X₁₈ is Phe or Leu;

X₁₉ is Trp, His, Gln or Pro;

X₂₀ is Leu or Ala;

X₂₁ is Trp or His;

X₂₂ is Pro or Trp;

X₂₃ is Ser, Ala, Pro or Gln;

X₂₄ is Ser, His, or Phe;

X₂₅ is Asp, Ser, Gln or Trp; and

X₂₆ is Phe, His, Ala or Asp,

wherein said binding peptide is displayed on the surface of said batereriophage.

4. A method for detecting human parvovirus B19 or B19-like polypeptide in a sample of blood or a blood fraction, comprising contacting said sample with a bacteriophage according to any one of claims 1, 2 or 3 and detecting whether binding has occurred between said bacteriophage and a human parvovirus B19 and/or a B19-like polypeptide.

5. A method for removing human parvovirus B19 or B19-like polypeptide from a solution containing it comprising:

(a) immobilizing a bacteriophage according to any one of claims 1, 2, or 3 on a solid support;

(b) contacting a solution containing human parvovirus B19 or B19-like polypeptide with said support; and

, thereafter,

(c) separating the solution from said support.

6. A method according to claim 5, wherein said solid support is a chromatographic matrix.

2. A recombinant bacteriophage expressing exogenous DNA encoding a parvovirus binding peptide having an amino acid sequence selected from the group consisting of:

Phe-Phe-Cys-Gly-Phe-Trp-His-Asp-Cys-His-Pro (SEQ ID NO;

     4);

Phe-Ser-Cys-Leu-Trp-Phe-Pro-Phe-Cys-Pro-Asp (SEQ ID NO;

     5);

Phe-Phe-Cys-Ala-Leu-Trp-Pro-Ser-Cys-His-His (SEQ ID NO;

     6);

Leu-Phe-Cys-His-Phe-Trp-Tyr-Asn-Cys-Asp-Phe (SEQ ID NO;

     7);

Leu-Phe-Cys-Ser-Phe-Trp-Tyr-Asn-Cys-Asp-Ala (SEQ ID NO;

     8);

Leu-Phe-Cys-Ser-Phe-Trp-Tyr-Asn-Cys-Asp-Asp (SEQ ID NO;

     9);

Leu-Phe-Cys-Arg-Phe-Trp-Tyr-Asn-Cys-Ser-Ala (SEQ ID NO;

     10);

Phe-Phe-Cys-Gln-Tyr-Trp-Tyr-Asn-Cys-Asp (SEQ ID NO;

     11);

Phe-Cys-Arg-Phe-Trp-Tyr-Gly-Cys-His-Pro (SEQ ID NO;

     12);

Phe-Phe-Cys-Ser-Phe-Trp-His-Gly-Gly-Cys-Asp-Asp (SEQ ID NO;

     13);

Ala-Phe-Cys-His-Phe-Trp-Phe-His-Gly-Cys-Asp-Asp (SEQ ID NO;

     14);

Ala-Phe-Cys-Trp-Lys-Trp-Pro-Gly-Asn-Cys-Lys-His (SEQ ID NO;

     15);

His-Phe-Cys-His-Phe-Trp-Phe-Gly-Gly-Cys-Pro-His (SEQ ID NO;

     16);

Phe-Cys-Trp-Leu-Trp-Pro-Ser-Ser-Asp-Cys-Phe (SEQ ID NO;

     17);

Phe-Cys-Trp-Leu-Trp-Pro-Ala-His-Ser-Cys-His (SEQ ID NO;

     18);

Phe-Cys-His-Leu-Trp-Trp-Pro-Phe-Gln-Cys-Ala (SEQ ID NO;

     19);

Phe-Cys-Gln-Leu-Trp-Trp-Pro-Phe-Gln-Cys-Ala (SEQ ID NO;

     20); and

Leu-Cys-Pro-Ala-His-Trp-Gln-Phe-Trp-Cys-Asp (SEQ ID NO;

     21),

wherein said binding peptide is displayed on the surface of said bacteriophage.

3. A recombinant bacteriophage expressing exogenous DNA encoding a parvovirus binding peptide having an amino acid sequence selected from the group consisting of:

Ala-Glu-Gly-Thr-Gly-Asp-Phe-Phe-Cys-Ser-Phe-Trp-His-Gly-Gly-Cys-Asp-Asp-Asp-Pro-Gly-Pro-Glu-Gly-Gly-Gly-Ser (SEQ ID NO;

     22); and

Ala-Glu-Gly-Thr-Gly-Asp-Phe-Cys-Trp-Leu-Trp-Pro-Ala-His-Ser-Cys-His-Asp-Pro-Gly-Pro-Glu-Gly-Gly-Gly-Ser (SEQ ID NO;

     23);

wherein said binding peptide is displayed on the surface of said bacteriophage.

7. A synthetic oligonucleotide encoding a parvovirus binding peptide, said binding peptide comprising an amino acid sequence selected from the group consisting of

I. X₁-X₂-Cys-X₃-X₄-X₅-X₆-X₇-Cys-X₈-X₉ (SEQ ID NO:

1), wherein

X₁ is Phe or Leu or is not present;

X₂ is Phe or Ser;

X₃ is Arg, Gln, Ser, His, Ala, Leu, or Gly;

X₄ is Phe, Tyr, Leu, or Trp;

X₅ is Trp or Phe;

X₆ is Tyr, Pro, or His;

X₇ is Gly, Asn, Ser, Phe, or Asp;

X₈ is His, Asp, Ser or Pro;

X₉ is Pro, Ala, Phe, His, or Asp or is not present,

II. X₁₀-Phe-Cys-X₁₁-X₁₂-Trp-X₁₃-X₁₄-X₁₅-Cys-X₁₆-X₁₇ (SEQ ID NO;

     2), wherein

X₁₀ is His, Ala, or Phe;

X₁₁ is His, Trp, or Ser;

X₁₂ is Phe or Leu;

X₁₃ is Phe, Pro, or His;

X₁₄ is Gly or His;

X₁₅ is Gly or Asn;

X₁₆ Pro, Leu, or Asp;

X₁₇ is His or Asp,

III. X₁₈-Cys-X₁₉-X₂₀-X₂₁-X₂₂-X₂₃-X₂₄-X₂₅Cys-X₂₆ (SEQ ID NO;

     3), wherein

X₁₈ is Phe or Leu;

X₁₉ is Trp, His, Gln or Pro;

X₂₀ is Leu or Ala;

X₂₁ is Trp or His;

X₂₂ is Pro or Trp;

X₂₃ is Ser, Ala, Pro or Gln;

X₂₄ is Ser, His, or Phe;

X₂₅ is Asp, Ser, Gln or Trp; and

X₂₆ is Phe, His, Ala or Asp.

8. The oligonucleotide according to claim 7, wherein said oligonucleotide encodes a binding peptide comprising an amino acid sequence selected from the group consisting of:

Phe-Phe-Cys-Gly-Phe-Trp-His-Asp-Cys-His-Pro (SEQ ID NO;

     4);

Phe-Ser-Cys-Leu-Trp-Phe-Pro-Phe-Cys-Pro-Asp (SEQ ID NO;

     5);

Phe-Phe-Cys-Ala-Leu-Trp-Pro-Ser-Cys-His-His (SEQ ID NO;

     6);

Leu-Phe-Cys-His-Phe-Trp-Tyr-Asn-Cys-Asp-Phe (SEQ ID NO;

     7);

Leu-Phe-Cys-Ser-Phe-Trp-Tyr-Asn-Cys-Asp-Ala (SEQ ID NO;

     8);

Leu-Phe-Cys-Ser-Phe-Trp-Tyr-Asn-Cys-Asp-Asp (SEQ ID NO;

     9);

Leu-Phe-Cys-Arg-Phe-Trp-Tyr-Asn-Cys-Ser-Ala (SEQ ID NO;

     10);

Phe-Phe-Cys-Gln-Tyr-Trp-Tyr-Asn-Cys-Asp (SEQ ID NO;

     11);

Phe-Cys-Arg-Phe-Trp-Tyr-Gly-Cys-His-Pro (SEQ ID NO;

     12);

Phe-Phe-Cys-Ser-Phe-Trp-His-Gly-Gly-Cys-Asp-Asp (SEQ ID NO;

     13);

Ala-Phe-Cys-His-Phe-Trp-Phe-His-Gly-Cys-Asp-Asp (SEQ ID NO;

     14);

Ala-Phe-Cys-Trp-Lys-Trp-Pro-Gly-Asn-Cys-Lys-His (SEQ ID NO;

     15);

His-Phe-Cys-His-Phe-Trp-Phe-Gly-Gly-Cys-Pro-His (SEQ ID NO;

     16);

Phe-Cys-Trp-Leu-Trp-Pro-Ser-Ser-Asp-Cys-Phe (SEQ ID NO;

     17);

Phe-Cys-Trp-Leu-Trp-Pro-Ala-His-Ser-Cys-His (SEQ ID NO;

     18);

Phe-Cys-His-Leu-Trp-Trp-Pro-Phe-Gln-Cys-Ala (SEQ ID NO;

     19);

Phe-Cys-Gln-Leu-Trp-Trp-Pro-Phe-Gln-Cys-Ala (SEQ ID NO;

     20); and

Leu-Cys-Pro-Ala-His-Trp-Gln-Phe-Trp-Cys-Asp (SEQ ID NO;

     21).

9. The oligonucleotide according to claim 7, wherein said oligonucleotide encodes a binding peptide comprising an amino acid sequence selected from the group consisting of:

Ala-Glu-Gly-Thr-Gly-Asp-Phe-Phe-Cys-Ser-Phe-Trp-His-Gly-Gly-Cys-Asp-Asp-Asp-Pro-Gly-Pro-Glu-Gly-Gly-Gly-Ser (SEQ ID NO;

     22); and

Ala-Glu-Gly-Thr-Gly-Asp-Phe-Cys-Trp-Leu-Trp-Pro-Ala-His-Ser-Cys-His-Asp-Pro-Gly-Pro-Glu-Gly-Gly-Gly-Ser (SEQ ID NO;

     23).

10. A recombinant expression vector comprising a polynucleotide according to any one of claims 7-9.

11. A recombinant host cell transformed with the expression vector according to claim 10.

12. A method of producing a human parvovirus binding peptide, comprising the steps:

(a) culturing a host cell according to claim 11 under conditions that promote expression of said binding peptide; and

(b) isolating said binding peptide from said host cell.