# US 2010/0129388 A1 · Protective proteins of s. agalactiae, combinations thereof and methods of using the same

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- Number: US 2010/0129388 A1
- Document ID: US20100129388A1
- Status: Published application
- Kind: A1
- Application number: 12/522,636
- Filed: 2008-01-10
- Published: 2010-05-27
- Earliest claimed: 2007-01-12
- Assignees: Intercell AG
- Inventors: Beatrice Senn; Eszter Nagy; Andreas Meinke; Alexander Von Gabain; Barbara Maierhofer; Ulrike Stierschneider; Manfred Berger; Christina Neubauer; Katherine Cohen
- Web page: https://patentgrep.com/patent/US20100129388A1

## CPC

- A61K 39/092: {Streptococcus}
- A61P 31/04: Antibacterial agents
- C07K 14/315: from Streptococcus (G), e.g. Enterococci
- C07K 16/1275: Streptococcus (G)
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- A61K 2039/505: {comprising antibodies}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The invention relates to a composition comprising at least two protective proteins against Streptococcus agalactiae (S. agalactiae) or functionally active variant thereof; a protective peptide against S. agalactiae; one or more nucleic acid(s) encoding the at least two proteins and/or the protective peptide; a method of producing the composition; a pharmaceutical composition, especially a vaccine, comprising the composition and/or at least one protective peptide; methods for producing antibodies; a mixture of antibodies against the at least two proteins of the composition; the use of the composition and/or at least one protective peptide and/or one or more nucleic acid(s) for the manufacture of a medicament for the immunization or treatment of a subject; methods of diagnosing a S. agalactiae infection; a method for identifying a ligand capable of binding the composition and/or at least one protective peptide; and the use of the composition and/or at least one protective peptide for the isolation and/or purification and/or identification of an interaction partner of the composition and/or peptide.

## Claims

### Claim 1

A composition comprising at least two proteins selected from the group consisting of
i) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 1 (gbs0233p) or functionally active variant thereof;
ii) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 2 (gbs1087p) or functionally active variant thereof;
iii) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 3 (gbs1309p) or functionally active variant thereof;
iv) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 4 (gbs1477p) or functionally active variant thereof;
v) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 5 (gbs1478p) or functionally active variant thereof; and
vi) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 6 (gbs2018p) or functionally active variant thereof.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the composition comprises at least three proteins selected from the group consisting of subgroup i) to vi).

### Claim 3 (depends on 1, 2)

The composition of claim 1 or 2, wherein the composition comprises at least four proteins selected from the group consisting of subgroup i) to vi).

### Claim 4 (depends on 1, 2, 3)

The composition of any of claims 1 to 3, wherein the at least two, three or four proteins are selected from different subgroups i) to vi).

### Claim 5 (depends on 1, 2, 3, 4)

The composition of any of claims 1 to 4, wherein at least two of the proteins are selected from one of the subgroups i) to vi).

### Claim 6 (depends on 1, 2, 3, 4, 5)

The composition of any of claims 1 to 5 comprising
at least one protein of subgroup iv);
at least one protein of subgroup vi);
at least one protein of subgroup iv) and at least one protein of subgroup vi);
at least one protein of subgroup iv), at least one protein of subgroup vi) and at least one protein of subgroup ii); or
at least one protein of subgroup iv), at least one protein of subgroup vi), at least one protein of subgroup ii) and at least one protein of subgroup v).

### Claim 7 (depends on 1, 2, 3, 4, 5, 6)

The composition of any of claims 1 to 6, wherein one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 4 (gbs1477p) or functionally active variant thereof, preferably protective peptide of SEQ ID NO: 4 (gbs1477p) or a naturally occurring variant thereof.

### Claim 8 (depends on 1, 2, 3, 4, 5, 6, 7)

The composition of any of claims 1 to 7, wherein one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 6 (gbs2018p) or functionally active variant thereof, preferably protective peptide of SEQ ID NO: 6 (gbs2018p) or a naturally occurring functionally active variant thereof.

### Claim 9 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The composition of any of claims 1 to 8, wherein one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 2 (gbs1087p) or functionally active variant thereof, preferably protective peptide of SEQ ID NO: 2 (gbs1087p) or a naturally occurring functionally active variant thereof.

### Claim 10 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9)

The composition of any of claims 1 to 9, wherein one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 5 (gbs1478p) or functionally active variant thereof, preferably protective peptide of SEQ ID NO: 5 (gbs1478p) or a naturally occurring functionally active variant thereof.

### Claim 11 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10)

The composition of any of claims 1 to 10, wherein one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 1 (gbs0233p) or functionally active variant thereof, preferably protective peptide of SEQ ID NO: 1 (gbs0233p) or a naturally occurring functionally active variant thereof.

### Claim 12 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11)

The composition of any of claims 1 to 11, wherein one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 3 (gbs1309p) or functionally active variant thereof, preferably protective peptide of SEQ ID NO: 3 (gbs1309p) or a naturally occurring functionally active variant thereof.

### Claim 13 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)

The composition of any of claims 1 to 12, wherein the at least two proteins encompass:
the protective peptide of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p);
the protective peptide of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p) and the protective peptide of SEQ ID NO: 2 (gbs1087p); or
the protective peptide of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p) and the protective peptide of SEQ ID NO: 2 (gbs1087p) and the protective peptide of SEQ ID NO: 5 (gbs1478p).

### Claim 14 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)

The composition of any of claims 1 to 13, wherein two or more proteins of the at least two proteins are combined into at least one fusion protein.

### Claim 15 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)

The composition of any of claims 1 to 14, wherein the functionally active variant
a) is a functionally active fragment of the protective peptide, the functionally active fragment comprising at least 50% of the sequence of the protective peptide, preferably at least 70%, more preferably at least 80%, still more preferably at least 90%, even more preferably at least 95% and most preferably at least 97%, 98% or 99%;
b) is derived from the protective peptide by at least one amino acid substitution, addition and/or deletion, wherein the functionally active variant has a sequence identity to the protective peptide or to the functionally active fragment as defined in a) of at least 40%, preferably at least 60%, more preferably at least 75%, still more preferably at least 90%, even more preferably at least 95% and most preferably at least 97%, 98% or 99%; and/or
c) consists of the protective peptide or a functionally active variant thereof and additionally at least one amino acid heterologous to the protective peptide,
preferably wherein the functionally active variant is derived from or identical to any of the naturally occurring variants of any of the sequences of SEQ ID NO: 55 to 60, 67 to 72, 79 to 84, 91 to 132, 175 to 179, 185 to 203, 223 to 234, and 235 to 425.

### Claim 16

A protective peptide consisting of the amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 55 to 57, 59, 60, 68, 69, 71, 72, 79 to 84, 91 to 132, 175 to 179, 185 to 203, 223 to 234, and 235 to 425.

### Claim 17 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

One or more nucleic acid(s) encoding the at least two proteins comprised in of the composition according to any of claims 1 to 15 and/or any of the protective peptides according to claim 16.

### Claim 18 (depends on 17)

The one or more nucleic acid(s) of claim 17, comprising or consisting of at least one nucleic acid sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 61 to 66, SEQ ID NO: 73 to 78, SEQ ID NO: 85 to 90, SEQ ID NO: 133 to 174, SEQ ID NO: 180 to 184 and. SEQ ID NO: 204 to 222.

### Claim 19 (depends on 17, 18)

The one or more nucleic acid(s) of claims 17 to 18, wherein the nucleic acid(s) is/are located in a vector or a cell other than S. agalactiae.

### Claim 20 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

A method of producing the composition of any of claims 1 to 15 or the protective peptide of claim 16, comprising
(a) introducing the one or more nucleic acids into a host cell;
(b) expressing the protein(s) and/or peptide(s) encoded by the nucleic acid by culturing the host cell under conditions conducive to the expression of the protein(s) and/or peptide(s); and
(c) collecting and/or isolating the expressed protein(s) and/or peptide(s) of step (b).

### Claim 21 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

A pharmaceutical composition, especially a vaccine, comprising
(i) the composition according to any of claims 1 to 15 and/or at least one protective peptide according to claim 16; and
(ii) optionally a pharmaceutically acceptable carrier or excipient.

### Claim 22 (depends on 17, 18, 19)

A pharmaceutical composition comprising
(i) the one or more nucleic acid(s) according to any of claims 17 to 19 or one or more nucleic acid(s) complementary thereto, and
(ii) optionally a pharmaceutically acceptable carrier or excipient.

### Claim 23 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

A method for producing antibodies, characterized by the following steps:
(a) administering an effective amount of the composition according to any of claims 1 to 15 and/or at least one protective peptide according to claim 16 to an animal; and
(b) isolating the antibodies produced by the animal in response to the administration of step (a) from the animal.

### Claim 24 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

A method for producing antibodies, characterized by the following steps:
(a) contacting a B cell with an effective amount of the composition according to any of claims 1 to 15 and/or at least one protective peptide according to claim 16;
(b) fusing the B cell of step (a) with a myeloma cell to obtain a hybridoma cell; and
(c) isolating the antibodies produced by the cultivated hybridoma cell.

### Claim 25 (depends on 23, 24)

The method of claim 23 or 24, wherein the isolated antibodies are additionally purified.

### Claim 26 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

A mixture of antibodies against the at least two proteins of the composition of any of claims 1 to 15 and/or against the at least one protective peptide according to claim 16.

### Claim 27 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)

Use of the composition according to any of claims 1 to 15 and/or at least one protective peptide according to claim 16 and/or one or more the nucleic acid(s) according to any of claims 17 to 19 for the manufacture of a medicament for the immunization or treatment of a subject, preferably against S. agalactiae, more preferably against pneumonia, septicemia, meningitis, fever, vomiting, poor feeding, irritability, urinary tract infection and/or vaginal infection caused by S. agalactiae.

### Claim 28 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

A method of diagnosing a S. agalactiae infection comprising the steps of:
(a) contacting a sample obtained from a subject with the composition according to any of claims 1 to 15 and/or at least one protective peptide according to claim 16; and
(b) detecting the presence of an antibody against the protective peptide, the functionally active variant and/or the composition in the sample,
wherein the presence of the antibody is indicative for the S. agalactiae infection.

### Claim 29 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 26)

A method of diagnosing a S. agalactiae infection comprising the steps of:
(a) contacting a sample obtained from a subject with the mixture of antibodies according to claim 26; and
(b) detecting the presence of the at least two proteins of the composition of any of claims 1 to 15 and/or of the at least one protective peptide according to claim 16 in the sample,
wherein the presence of the at least two proteins is indicative for the S. agalactiae infection.

### Claim 30 (depends on 17, 18, 19)

A method for diagnosing an infection with S. agalactiae comprising the steps of:
(a) contacting a sample obtained from a subject with a primer and/or a probe specific for the one or more nucleic acid(s) of any of claims 17 to 19; and
(b) detecting the presence of one or more nucleic acid(s) of any of claims 17 to 19 in the sample,
wherein the presence of the one or more nucleic acid(s) is indicative for the S. agalactiae infection.

### Claim 31 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

A method for identifying a ligand capable of binding the composition according to any of claims 1 to 15 and/or at least one protective peptide according to claim 16 comprising:
(a) providing a test system comprising the peptide and/or composition,
(b) contacting the test system with a test compound, and
(c) detecting a signal generated in response to the binding of the test compound to the peptide and/or composition.

### Claim 32 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)

Use of the composition according to any of claims 1 to 15 and/or at least one protective peptide according to claim 16 for the isolation and/or purification and/or identification of an interaction partner of the composition and/or the peptide.