Lonza Biologics Plc

Pfizer Corporation

The University of Manchester

Robert Young

James Rance

Marc Feary

Michael J Agostino

Mark A Moffat

Tom Payne

Baohong Zhang

Lin Zhang

Ian Kinloch

Tanya Jeane Knight

Christopher M Smales

Lei Gong

Mark C Smales

James D Budge

Elizabeth C Sayer

James Budge

Stephan Kalwy

Robert Gay

C Mark Smales

Christopher Mark Smales

The present invention relates to an expression system for the heterologous expression of a nucleic acid sequence of interest in a mammalian cell, the system comprising: (i) a first genetic entity, comprising: a nucleic acid sequence encoding a functional Epstein Barr virus nuclear antigen 1 (EBNA-1), the nucleic acid sequence being operably linked to regulatory elements that allow for expression of the nucleic acid sequence encoding a functional EBNA-1; (ii) a second genetic entity, comprising: a nucleic acid sequence encoding a functional nucleoside diphosphate kinase A (NDPK-A), the nucleic acid sequence being operably linked to regulatory elements that allow for expression of the nucleic acid sequence encoding a functional NDPK-A; (iii) a third genetic entity, comprising: the nucleic acid sequence of interest being operably linked to regulatory elements that allow for expression of the nucleic acid sequence of interest; and (iv) a four genetic entity, comprising: the Epstein Barr virus OriP sequence or one or more subsequences thereof, wherein the one or more subsequences comprise at least the 'Family of Repeats' DNA-binding site for EBNA-1 and the 'Dyad Symmetry' DNA-binding site for EBNA-1. The present invention also relates to corresponding mammalian host cells and methods for expressing a nucleic acid sequence of interest by means of such expression system.

Mammalian expression system

The present invention relates to stable and high-producing site-specific integration (SSI) host cells, e. g. Chinese hamster ovary (CHO)-derived host cells, methods to produce and to use them.

Site-specific integration

The present invention relates to novel nanocomposite materials, methods of making nanocomposites and uses of nanocomposite materials. In particular, the invention relates to composite materials which contain two-dimensional materials (e.g. graphene) in multi-layer form i.e. in a form which has a number of atomic layers. The properties of a composite material containing two-dimensional material is in multi-layer from are shown to be superior to those which contain the two-dimensional material in monolayer form.

Graphene composites

Methods for generating mammalian cells characterized by an increased concentration of at least one of miR-15, miR-16, and miR-34 for producing proteins at an industrial scale are provided. Methods for using the mammalian cells, in particular for the production of proteins of interest, are also provided.

Means and methods for the generation of mammalian producer cells for the production of recombinant proteins

The present invention relates to expression vectors for the heterologous expression of a nucleic acid sequence of interest in mammalian cells, the vectors comprising a chimeric promoter regulatory sequence being operably linked to a nucleic acid sequence to be expressed, wherein the chimeric promoter regulatory sequence comprises a cytomegalovirus promoter sequence derived from murine cytomegalovirus or from human cytomegalovirus and being operably linked to the transcriptional start site of the nucleic acid sequence to be expressed; and a cytomegalovirus upstream region and/or enhancer sequence derived from human and/or the simian cytomegalovirus, wherein the upstream region and/or enhancer sequence is located 5' of and operably linked to the murine or the human promoter sequence, and wherein the chimeric promoter regulatory sequence comprises sequence elements being derived from at least two of the group consisting of murine cytomegalovirus, human cytomegalovirus and simian cytomegalovirus. In particular embodiments, the chimeric promoter regulatory sequence comprises sequence elements derived from the murine or the human cytomegalovirus IE1 promoter and from the human and/or the simian cytomegalovirus IE1 region. The invention also relates to mammalian host cells transfected with such expression vectors, a method for heterologous expression of a nucleic acid sequence in a mammalian host cell by employing such expression vectors, and the use of such expression vectors for the heterologous expression of a nucleic acid sequence.

Expression vectors comprising chimeric cytomegalovirus promoter and enhancer sequences

The present invention relates to a mammalian cell based expression and secretion system and the expression and secretion of recombinant proteins by using secretory signal peptides. The present invention also relates to an expression cassette useful for the secretion of a heterologous gene from a mammalian cell, in particular a CHO cell. The present invention is also directed to a method of secreting a heterologous protein from mammalian cells such as CHO cells.

Mammalian expression vector with a highly efficient secretory signal sequence

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London, United Kingdom

Robert Young