Glaxosmithkline Biologicals Sa

University of Alberta

Amirreza Faridmoayer

Michael T Kowarik

Fabio Serventi

Stefan Jochen Kemmler

Rainer Follador

Gerald Johann Posch

Gerd Martin Lipowsky

Sabina Marietta Gerber

Michael L Wetter

Michael Wacker

Martin Edward Braun

Mario Feldman

Markus Mueller

A process for engineering a host cell comprising the steps of a) integrating a first polynucleotide cassette including a first selection marker flanked by a first pair of recombination sites; b) removing the first selection marker by the action of a recombinase which recognises the first pair of recombination sites; c) integrating a second polynucleotide cassette including a second selection marker flanked by a second pair of recombination sites; and d) removing the second selection marker by the action of a recombinase which recognises the second pair of recombination sites. The first and second pairs of recombination sites have identical nucleic acid sequences within each pair, sharing 90-98% nucleic acid sequence identity. Also disclosed is a host cell genome polynucleotide comprising two recombinantly engineered regions, each adjacent to a single recombination site sharing 90-98% identity with each other and any additional recombination sites present in the host cell genome polynucleotide.

Process for the manipulation of nucleic acids

The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharides comprising one or more repeat unit(s) →4)-β-D-Glcp-(1→4)-[Gro-(2→P→3)]-β-D-Galp-(1→4)-β-L-Rhap-(1→. Also provided are conjugates comprising a →4)-β-D-Glcp-(1→4)-[Gro-(2→P→3)]-β-D-Galp-(1→4)-β-L-Rhap-(1→, immunogenic compositions, vaccines and their use in preventing or treating infection by

Immunogenic composition

The present invention discloses a process for engineering a host cell comprising the steps of; a) integrating a first polynucleotide cassette including a first selection marker flanked by a first pair of recombination sites; b) removing the first selection marker by the action of a recombinase which recognises the first pair of recombination sites; c) integrating a second polynucleotide cassette including a second selection marker flanked by a second pair of recombination sites; and d) removing the second selection marker by the action of a recombinase which recognises the second pair of recombination sites. Also disclosed is a host cell genome polynucleotide comprising a first recombinantly engineered region and a second recombinantly engineered region, wherein a first single recombination site is adjacent to the first recombinantly engineered region, and a second single recombination site is adjacent to the second recombinantly engineered region.

The present invention discloses modifiedClfA proteins that contain glycosylation site consensus sequences. The invention also discloses a conjugate comprising a modified ClfA protein and an antigen (for example asaccharide antigen), wherein the antigen is linked (either directly or through a linker) to an amino acid residue of the modified ClfA protein.

The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharide comprising one or more repeat unit(s) &#x2192;4)-&#x3b2;-D-Glcp-(1&#x2192;3)-[[&#x3b1;-L-Rhap-(1&#x2192;2)]-[Gro-(2&#x2192;P&#x2192;3)]-&#x3b2;-D-Galp-(1&#x2192;4)]-&#x3b2;-L-Rhap-(1&#x2192;. Also provided are conjugates comprising a &#x2192;4)-&#x3b2;-D-Glcp-(1&#x2192;3)-[[&#x3b1;-L-Rhap-(1&#x2192;2)]-[Gro-(2&#x2192;P&#x2192;3)]-&#x3b2;-D-Galp-(1&#x2192;4)]-&#x3b2;-L-Rhap-(1&#x2192;, immunogenic compositions, vaccines and their use in preventing or treating infection by

The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharides comprising one or more repeat unit(s) &#x2192;4)-&#x3b2;-D-Glcp-(1&#x2192;4)-[Gro-(2&#x2192;P&#x2192;3)]-&#x3b2;-D-Galp-(1&#x2192;4)-&#x3b2;-L-Rhap-(1&#x2192;. Also provided are conjugates comprising a &#x2192;4)-&#x3b2;-D-Glcp-(1&#x2192;4)-[Gro-(2&#x2192;P&#x2192;3)]-&#x3b2;-D-Galp-(1&#x2192;4)-&#x3b2;-L-Rhap-(1&#x2192;, immunogenic compositions, vaccines and their use in preventing or treating infection by

The present invention discloses modifiedHIa proteins which show reduced tendency to aggregate, improving protein stability and yield. Said modified HIa proteins optionally also contain glycosylation site consensus sequences. The invention also discloses a conjugate comprising a modified HIa protein and an antigen (for example asaccharide antigen), wherein the antigen is linked to an amino acid residue of the modified HIa protein.

Provided herein are host cells capable of producing hybrid oligosaccharides and polysaccharides, wherein said hybrid oligosaccharides and polysaccharides do not comprise a hexose at the reducing end of their first repeat unit. Also provided herein are hybrid oligosaccharides or polysaccharides and bioconjugates which can be produced by the host cells described herein, wherein said bioconjugates comprise a carrier protein linked to a hybrid oligosaccharide or polysaccharide that does not comprise a hexose at the reducing end of its first repeat unit.

Modified host cells and hybrid oligosaccharides for use in bioconjugate production

Provided herein are prokaryotic cells proficient to produce glycoconjugates in vivo, as well as methods for generating these cell and methods of using these cells to produce glycoconjugates. The compositions of the mentioned glycoconjugates as well as their different uses are also included.

Production of recombinant vaccine inby enzymatic conjugation

Described herein are methods and systems for O-glycosylating proteins in vivo or in vitro in any prokaryotic organism. In these methods and systems, DNA comprising a gene that produces a PglL-like oligosaccharyltransferase and DNA comprising a gene that produces a protein to be O-glycosylated are used. The PglL-like oligosaccharyltransferase facilitates the covalent attachment of the glycan to the protein to produce the O-glycosylated protein. The methods and systems described herein provide an approach for the design and production of new vaccines and therapeutic agents for the treatment of various diseases.

Growing community of inventors

Schlieren, Switzerland

Amirreza Faridmoayer