Glaxosmithkline Biologicals Sa

Novartis Ag

Chiron S.p.a.

Other

Novartis Vaccines and Diagnostics Srl

Paolo Costantino

Guido Grandi

Francesco Berti

Maria Rosaria Romano

Rino Rappuoli

Mario Contorni

John Telford

Roberto Adamo

Laura Serino

Giuseppe Del Giudice

Domenico Maione

Francesco Norelli

Francesca Micoli

Roberto Olivieri

Oretta Finco

Giulia Bazzocchi

Concetta Cicala

Silvia Fontani

Anna Kabanova

Aldo Giannozzi

Angela Bardotti

Stefan Oscarson

Giovanni Averani

Costante Ceccarini

Marta Tontini

Peter Teodorovic

Stefano Viti

Sandro D'Ascenzi

Pierino Parente

Bruno Galletti

Giulia Brogioni

Maria Romano

Maria Rosario Romano

Concetta Maria Cicala

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

Precipitated bacterial capsular polysaccharides can be efficiently re-solubilised using alcohols as solvents. The invention provides a process for purifying a bacterial capsular polysaccharide, comprising the steps of (a) precipitation of said polysaccharide, followed by (b) solubilisation of the precipitated polysaccharide using ethanol. CTAB can be used for step (a). The material obtained, preferably following hydrolysis and sizing, can be conjugated to a carrier protein and formulated as a vaccine. Also, in vaccines comprising saccharides from both serogroups A and C, the invention provides that the ratio (w/w) of Men A saccharide:MenC saccharide is &#x3e;1.

Capsular polysaccharide solubilisation and combination vaccines

An injectable immunogenic composition comprising capsular saccharides from at least two of serogroups A, C, W135 and Y of, wherein said capsular saccharides are conjugated to carrier protein(s) and/or are oligosaccharides, and wherein (i) the composition comprises &#x3c;50 &#x3bc;g meningococcal saccharide per dose, and/or (ii) the composition further comprises an antigen from one or more of: (a) serogroup B; (b)type B; and/or (c). Saccharide antigens in the compositions are generally conjugated to a carrier.

Injectable vaccines against multiple meningococcal serogroups

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

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 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.

An injectable immunogenic composition comprising capsular saccharides from at least two of serogroups A, C, W135 and Y of, wherein said capsular saccharides are conjugated to carrier protein(s) and/or are oligosaccharides, and wherein (i) the composition comprises &#x3c;50 &#x3bc;g meningococcal saccharide per dose, and/or (ii) the composition further comprises an antigen from one or more of (a) serogroup B; (b)type B; and/or (c). Saccharide antigens in the compositions are generally conjugated to a carrier.

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.

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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

Precipitated bacterial capsular polysaccharides can be efficiently re-solubilized using alcohols as solvents. The invention provides a process for purifying a bacterial capsular polysaccharide, comprising the steps of (a) precipitation of said polysaccharide, followed by (b) solubilization of the precipitated polysaccharide using ethanol. CTAB can be used for step (a). The material obtained, preferably following hydrolysis and sizing, can be conjugated to a carrier protein and formulated as a vaccine. Also, in vaccines comprising saccharides from both serogroups A and C, the invention provides that the ratio (w/w) of MenA saccharide:MenC saccharide is &#x3e;1.

Carrier-induced epitopic suppression is of particular concern where multiple conjugates with the same carrier protein are administered simultaneously. To avoid the suppression, the invention minimizes the amount of unconjugated carrier protein in a vaccine. The invention provides a composition for immunizing a patient against a disease caused by, wherein (1) the composition comprises conjugates for at least two of the four meningococcal serogroups A, C, W135 and Y, where at least two of the conjugates have a common carrier protein; and (2) the composition includes the common carrier in an unconjugated form at less than 10 &#x3bc;g/ml.

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Siena, Italy

Paolo Costantino