Merck Sharp + Dohme Corp.

Merck + Co., Inc.

Other

Michael Albert Winters

Patrick McHugh

Emily Wen

Jian He

William J Smith

Chitrananda Abeygunawardana

Patrick L Ahl

Luwy Kavuka Musey

Akhilesh Bhambhani

Julie Marie Skinner

Jessica Sinacola

Morrisa C Jones

Matthew P Watson

Justin Stanbro

Christopher Jon Farrell

Daniel D Roth

Yadong Adam Cui

Michael J Kosinski

Russel Jackson Lander

John E MacNair

Jitin Bajaj

Francis Jeremiah Meacle

John M Williams

Joseph G Joyce

Janelle Konietzko

Sheng-Ching Wang

Cecilia Giovarelli

Hari Pujar

Thomas Svab

Catherine Lancaster

Jason C Murphy

David B Boyd

Adam Joel Kristopeit

Patrick Mchugh

John E Macnair

Patrick Leonard Ahl

Jessica R Sinacola

A method is described for producing a pneumococcal capsular polysaccharide protein conjugate in which one or more activated pneumococcal polysaccharides of particular pneumococcal serotypes and carrier protein are separately lyophilized, the separately lyophilized polysaccharides and carrier protein are separately reconstituted in an organic solvent, and the reconstituted polysaccharide and carrier protein are then combined together by Tee-mixing and conjugated together to produce polysaccharide carrier protein conjugates. A plurality of conjugates, each comprising polysaccharides of a particular serotype, may be used to produce multivalent pneumococcal immunogenic compositions having a combination of conjugates for use in vaccines.

Methods for producingcapsular polysaccharide carrier protein conjugates

The present invention provides a number of process improvements related to the conjugation of capsular polysaccharides fromto a carrier protein. These process are serotype specific and include acid hydrolysis, addition of sodium chloride to the reductive amination reaction, and addition of sucrose to dissolve polysaccharides. Polysaccharide-protein conjugates prepared using the processes of the invention can be included in multivalent pneumococcal conjugate vaccines.

Processes for the formulation of pneumococcal polysaccharides for conjugation to a carrier protein

A method is described for producing a pneumococcal capsular polysaccharide protein conjugate in which one or more activated pneumococcal polysaccharides of particular pneumococcal serotypes and carrier protein are separately lyophilized in the form of lyospheres of carrier protein and lyospheres of the one or more polysaccharides. A predetermined amount of carrier protein lyospheres and activated polysaccharide lyospheres are mixed together and the mixture reconstituted in an organic solvent to produce polysaccharide carrier protein conjugates. A plurality of conjugates, each comprising polysaccharides of a particular serotype, may be used to produce multivalent pneumococcal immunogenic compositions having a combination of conjugates for use in vaccines.

Methods for producing streptococcus pneumoniae capsular polysaccharide carrier protein conjugates from lyospheres

The invention is related to multivalent immunogenic compositions comprising more than onepolysaccharide protein conjugates, wherein each of the conjugates comprises a polysaccharide from anserotype conjugated to a carrier protein, wherein the serotypes ofare as defined herein. Also provided are methods for inducing a protective immune response in a human patient comprising administering the multivalent immunogenic compositions of the invention to the patient. The multivalent immunogenic compositions are useful for providing protection againstinfection and diseases caused by. The compositions of the invention are also useful as part of treatment regimes that provide complementary protection for patients that have been vaccinated with a multivalent vaccine indicated for the prevention of pneumococcal disease.

Compositions comprisingpolysaccharide-protein conjugates and methods of use thereof

The present invention provides methods for making polysaccharide-protein conjugates in which polysaccharides, typically from bacteria, are conjugated to a carrier protein by reductive amination under conditions which improve conjugation reaction consistency, increase consumption of protein during conjugation reaction, generate conjugates of higher molecular weight, and/or reduce the levels of free cyanide in the conjugate reaction product. The polysaccharide-protein conjugates obtained using these methods are useful for inclusion in multivalent vaccines.

Methods for making polysaccharide-protein conjugates

A method is described for reconstituting lyophilized mutant diphtheria toxin in dimethyl sulfoxide for use in producing pneumococcal capsular polysaccharide mutant diphtheria toxin conjugates.

Methods for providing a homogenous solution of lyophilized mutant diptheria toxin in dimethylsulfoxide

The present invention provides a method of producing a polysaccharide-protein conjugate with capsular polysaccharide fromserotype 19F conjugated to a carrier protein. The method includes a prolonged incubation step prior to filtration to remove free polysaccharide.

Methods for production of capsular polysaccharide protein conjugates fromserotype 19F

The invention is related to multivalent immunogenic compositions comprising more than onepolysaccharide protein conjugates, wherein each of the conjugates comprises a polysaccharide from anserotype conjugated to a carrier protein, wherein the serotypes ofare as defined herein. In some embodiments, at least one of the polysaccharide protein conjugates is formed by a conjugation reaction comprising an aprotic solvent. In further embodiments, each of the polysaccharide protein conjugates is formed by a conjugation reaction comprising an aprotic solvent. Also provided are methods for inducing a protective immune response in a human patient comprising administering the multivalent immunogenic compositions of the invention to the patient. The multivalent immunogenic compositions are useful for providing protection againstinfection and diseases caused by. The compositions of the invention are also useful as part of treatment regimes that provide complementary protection for patients that have been vaccinated with a multivalent vaccine indicated for the prevention of pneumococcal disease.

The present invention provides immunogenic compositions having one or more polysaccharide-protein conjugates in which polysaccharides obtained from bacterial capsules are conjugated to diphtheria toxin fragment B (DTFB) or a variant thereof. The immunogenic compositions may be multivalent pneumococcal polysaccharide conjugate compositions in which polysaccharides from one or moreserotypes are conjugated to diphtheria toxin fragment B (DTFB) and, optionally, additional polysaccharides from one or more differentserotypes are conjugated to one or more other carrier proteins.

Polysaccharide-protein conjugates utilizing diphtheria toxin fragment B as a carrier

The present invention provides immunogenic compositions having one or more polysaccharide-protein conjugates in which one or more polysaccharides frombacterial capsules are conjugated to a carrier protein in an aprotic solvent such as dimethylsulfoxide (DMSO). The present invention also provides methods for providing an enhanced immune response to a pneumococcal polysaccharide protein conjugate vaccine comprising administering to a human subject an immunogenic composition comprising polysaccharide-protein conjugates prepared in DMSO conditions.

Enhancing immunogenicity ofpolysaccharide-protein conjugates

Methods of isolating clinical-grade plasmid DNA from manufacturing processes, including large-scale fermentation regimes, are disclosed which encompass alternatives to two core unit operations common to plasmid DNA purification processes. The novel upstream and downstream purification processes disclosed herein provide for reduced production costs and increase process robustness. Either or both of the purification processes disclosed herein may be used in combination with additional purification steps known in the art that are associated with DNA plasmid purification technology.

Purification process for plasmid DNA

The present invention relates to a nonchromatographic-based process for the isolation of clinical grade plasmid DNA from bacterial cells. The exemplified methods described herein outline a scaleable, economically favorable protocol for the purification of clinical grade plasmid DNA fromwhich includes CTAB-based precipitation of DNA in combination with adsorption of impurities to calcium silicate.

Process for the scaleable purification of plasmid DNA

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Doylestown, PA, United States of America

Michael Albert Winters