Glaxosmithkline Biologicals Sa

Novartis Ag

Novartis Vaccines and Diagnostics Srl

Gsk Vaccines S.r.l.

Francesco Berti

Paolo Costantino

Maria Rosaria Romano

Roberto Adamo

Francesca Micoli

Qi-Ying Hu

Immaculada Margarit Y Ros

Francesco Norelli

Roberto Olivieri

Anna Kabanova

Giulia Bazzocchi

Concetta Cicala

Silvia Fontani

Guido Grandi

Mario Contorni

John Telford

Laura Serino

Giuseppe Del Giudice

Domenico Maione

Carl E Frasch

Cira Daniela Rinaudo

Oretta Finco

Angela Bardotti

Giovanni Averani

Valdemar Robert Janulczyk

Filippo Carboni

Chiara Toniolo

Massimo Bigio

Cinzia Bellucci

Elisa Danieli

Roberta Cozzi

Marta Tontini

Evita Balducci

Martin Allen

Giulia Brogioni

Concetta Maria Cicala

Maria Romano

Maria Rosario Romano

Pierino Parente

Bruno Galletti

This disclosure provides novel saccharide derivatives, conjugates, and methods for making the derivatives and conjugates.

Cycloalkyne derivatized saccharides

The present invention is directed towards conjugates comprising fragments of the capsular polysaccharide of Type III Group B(GBS). Suitable fragments may be produced synthetically or by depolymerisation of native polysaccharide.

Immunogenic compositions

The invention provides a method for releasing capsular polysaccharide fromtype 5 or type 8 cells, comprising the step of treating the cells with acid. The invention further provides a process for purifying capsular polysaccharide fromtype 5 or type 8 cells comprising this method. Other processing steps may be included in the process, such as enzymatic treatment, e.g. to remove nucleic acid, protein and/or peptidoglycan contaminants; diafiltration, e.g. to remove low molecular weight contaminants; anion exchange chromatography, e.g. to remove residual protein; and concentration. The invention also provides a composition comprising atype 5 or type 8 capsular polysaccharide, obtainable by any of the processes of the invention.

type 5 capsular saccharides

A process for purifying aGAS carbohydrate comprising a step of anionic exchange chromatography. The process provides a good yield of GAS carbohydrate. The saccharides of the invention have low levels of hyaluronic acid, protein and nucleic acid contamination.

Purification method

The invention provides a process for preparing a conjugate of atype 5 or type 8 capsular polysaccharide and a carrier molecule, comprising the steps of: (a) depolymerising the capsular polysaccharide, to give a polysaccharide fragment; (b) oxidising the fragment in order to introduce an aldehyde group into at least one saccharide residue in the fragment, to give an oxidised saccharide residue; and (c) coupling the oxidised saccharide residue to a carrier molecule via the aldehyde group, thereby giving the conjugate. The coupling in step (c) may be direct, or may be via a linker molecule. The invention also provides a conjugate obtained or obtainable by this process.

Conjugation oftype 5 and type 8 capsular polysaccharides

The invention provides a conjugate of aserogroup X capsular polysaccharide and a carrier molecule. The conjugate is typically made by (a) oxidising a primary hydroxyl group in the capsular polysaccharide, to give an oxidised polysaccharide with an aldehyde group; and (b) coupling the oxidised polysaccharide to a carrier molecule via the aldehyde group, thereby giving the conjugate. The conjugate may be part of an immunogenic composition. This composition may comprise one or more further antigens, particularly capsular polysaccharides from serogroups A, W135, C and Y and conjugated forms thereof. The composition may be in an aqueous formulation. The composition is useful as a vaccine, e.g. for raising an immune response in a mammal. The invention also provides processes for making the conjugate.

Meningococcus serogroup X conjugate

The invention provides a conjugate comprising an antigen and a carrier molecule, wherein the carrier molecule comprises a BP-2a antigen and a spb1 antigen. BP-2a and spb1 areantigens. The conjugate may be used in a method for raising an immune response in a mammal, the method comprising administering the conjugate to the mammal. Also provided are pharmaceutical compositions, particularly vaccines, comprising the conjugate.

Carrier molecule for antigens

Three conjugation methods for use with the capsular saccharide of. In the first method, reductive amination of oxidized sialic acid residue side chains is used, but the aldehyde groups are first aminated, and then the amine is coupled to a carrier via a linker. In the second method, sialic acid residues and/or N-acetyl-glucosamine residues are de-N-acetylated to give amine groups, and the amine groups are coupled to a carrier protein via a linker. In the third method, linkage is via galactose residues in the capsular saccharide rather than sialic acid residues, which can conveniently be achieved using galactose oxidase.

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A purification method for the capsular polysaccharide of type II GBS in which the capsular polysaccharide is filtered using a membrane with a cut-off of less than 30 kDa.

Purification of streptococcal capsular polysaccharide

The invention relates to bacterial mutants, particularly from, that secrete capsular polysaccharide and methods of purifying the secreted bacterial capsular polysaccharides from culture medium. The extracted polysaccharides are useful for producing vaccines comprising the polysaccharides alone or conjugated to proteins.

Purification of secreted polysaccharides from S. agalactiae

The invention provides a conjugate of aserogroup X capsular polysaccharide and a carrier molecule. The conjugate is typically made by (a) oxidizing a primary hydroxyl group in the capsular polysaccharide, to give an oxidized polysaccharide with an aldehyde group; and (b) coupling the oxidized polysaccharide to a carrier molecule via the aldehyde group, thereby giving the conjugate. The conjugate may be part of an immunogenic composition. This composition may comprise one or more further antigens, particularly capsular polysaccharides from serogroups A, W135, C and Y and conjugated forms thereof. The composition may be in an aqueous formulation. The composition is useful as a vaccine, e.g. for raising an immune response in a mammal. The invention also provides processes for making the conjugate.

The invention provides a process for preparing a conjugate of atype 8 capsular polysaccharide and a carrier molecule, comprising the steps of: (a) depolymerising the capsular polysaccharide, to give a polysaccharide fragment; (b) oxidising the fragment in order to introduce an aldehyde group into at least one saccharide residue in the fragment, to give an oxidised saccharide residue; and (c) coupling the oxidised saccharide residue to a carrier molecule via the aldehyde group, thereby giving the conjugate. The coupling in step (c) may be direct, or may be via a linker molecule. The invention also provides a conjugate obtained or obtainable by this process.

Capsular saccharides derived from serogroups W135 and Y ofhave altered levels of O-acetylation at the 7 and 9 positions of their sialic acid residues, and can be used to make immunogenic compositions. Relative to unmodified native saccharides, derivatives of the invention are preferentially selected during conjugation to carrier proteins, and conjugates of the derivatives show improved immunogenicity compared to native polysaccharides.

Hypo- and hyper-acetylated meningococcal capsular saccharides

Modified capsular saccharides comprising a blocking group at a hydroxyl group position on at least one of the monosaccharide units of the corresponding native capsular saccharide, wherein the blocking group is of the formula (Ia) or (Ib): &#x2014;OX&#x2014;Y (Ia) or &#x2014;O&#x2014;R(Ib) wherein X is C(O), S(O) or SO; Y is NRRor R; Ris Calkyl substituted with 1, 2 or 3 groups independently selected from hydroxyl, sulphydryl and amine; Ris H or Calkyl; and Ris Calkyl; processes for modifying a capsular saccharide with the blocking groups; saccharide-protein conjugates comprising the modified capsular saccharide; processes for making the saccharide-protein conjugates, pharmaceutical compositions comprising the modified capsular saccharides and/or saccharide-protein conjugates; and methods and uses of the same.

Modified saccharides

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Siena, Italy

Francesco Berti