Medimmune Limited

Suzanne J Gibson

Diane Hatton

David James

Daniel Ramsay Higazi

Nicholas J Bond

Mariana Alessia Sheriff

Sarah Milne

Adam J Brown

David C James

Daniel R Higazi

As demonstrated herein, when composite transcription factor binding sites do not function synergistically, mammalian promoters can be constructed according to simple design rules. Host-cell transcriptional machinery components were analyzed in silico to identify transcription factors with desired expression dynamics. Cognate binding sites were then comprehensively tested in homotypic and heterotypic architectures to assess modularity and determine the transcriptional activity exhibited by a single copy of each site. When elements were specifically selected to prevent combinatorial interactions, heterotypic promoter activities could be accurately modeled simply as a function of constituent binding site copy numbers. As binding site order, spacing, and orientation had minimal effect on promoter activity, blocks could be optimally combined and arranged in silico according to context-specific design-criteria. To demonstrate this, CHO cell promoters were created de-novo that exhibited designed activity levels and long-term expression stability in vitro.

In silico design of mammalian promoters with user-defined functionality

We discovered that recombinant antibody light chains having a murine secretory leader sequence and an SYE motif at the N-terminus are truncated during post-translational processing. This disclosure provides two protein engineering solutions: to alter the SYE amino acid sequence of the Lc N-terminus to other alternatives; or to change the secretory leader peptide sequence. We have shown that both of these solutions are effective for preventing N-terminal light chain truncation.

Prevention of N-terminal truncation in IgG light chains

Growing community of inventors

Cambridge, United Kingdom

Suzanne J Gibson