Glaxosmithkline Biologicals Sa

Gerd Martin Lipowsky

Michael T Kowarik

Amirreza Faridmoayer

Fabio Serventi

Stefan Jochen Kemmler

Rainer Follador

Gerald Johann Posch

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 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 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

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Schlieren, Switzerland

Gerd Martin Lipowsky