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Protein Optimization

Improved Expression 

Many multi-pass membrane proteins are difficult to over-express for research needs including antibody discovery and structural studies.

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Vaccinex combines mutagenesis with our proprietary vaccinia virus library technology to discover improved expression variants for these difficult targets.

Read our 2023 Nature Communications Paper "A Vaccinia-based system for directed evolution of GPCRs in mammalian cells"
cartoon illustrating poxvirus to sort libraries for GPCR mutations that improve expression. Poxvirus particles are shown in blue, infected cells are shown in gray with GPCRs in their outer membrane. Each cells has a different number of GPCRs in the membrane indicating variable expression. The cell with the most GPCRs in the membrane has a red dot in the GPCR that indicates a favorable mutation. Binding ligands are illustrated as bright green circles with a green binding domain extended out towards the GPCRs.

A Vaccinia virus library of 100 million unique mutant clones is generated and used to infect mammalian cells at 1 virus per cell.

Following infection, cDNAs encoded by vaccinia are expressed on the mammalian cell surface.

Libraries can be sorted using fluorescent binding molecules such as ligands to discover clones with expression levels higher than the native sequence.

Virus from the sorted cells are easily recovered for additional sorts or sequencing and characterization.

Cartoon illustrating a cDNA library of poxvirus infecting cells for sorting. Poxvirus are in blue, pink, purple and yellow indicating various cDNA sequences. Cells are in gray with transmembrane proteins in their membranes the same color as the poxvirus. The fluorescent binding molecule is indicated as green circles with a green binding domain extended out towards the protein at the cell surface.

Vaccinia library of cDNA from target (ex. tumor cell line)

Orpaned bining moleule with fluorescent detection

Virus recovered from sorted cells

Antigen Discovery

Our vaccinia library technology can also be applied to antigen discovery.
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We can generate a custom cDNA library from a target cell line and then sort the infected cells that are bound by the reagent of interest.

Learn more about our Vaccinia Virus Library Technology
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