Optimization of Complex Membrane Antigens

 

Vaccinia Display to Optimize Expression of Multi-pass Membrane Proteins

Many multi-pass membrane proteins have intrinsically low levels of expression which presents challenges to antibody discovery. Vaccinex can improve expression of these difficult targets by utilizing mutagenesis with our viral library technology.

Following infection, cDNA’s encoded by vaccinia are expressed as though native to the infected cell. Libraries of millions of variants can be screened in a fully mammalian system to discover clones with higher expression levels that maintain native antigen properties.

Protein-Optimization-1

 

Vaccinia expression of mutant library

  • Every cell expresses one mutant
  • Infected cells are stained for flow cytometry using labeled ligand or specific antibody
  • Cells with higher expression of the target are sorted
  • Recover virus from sorted cells by simple freeze/thaw
  • Confirm improved expression by flow cytometry
  • Deconvolute to identify individual mutants

 

Protein-Optimization-2