How to Improve 2A Linker Cleavage Efficiency

Here are some strategies to increase 2A linker cleavage efficiency for monoclonal antibody production:

Optimize the 2A peptide sequence: Different 2A peptides exhibit varying cleavage efficiencies. T2A with a GSG linker has shown the highest efficiency in some studies.
Add a furin recognition sequence: Placing a furin recognition sequence before the 2A peptide can help remove 2A residues from the upstream gene.
Insert GSG or SGS linkers: Adding GSG or SGS linkers immediately preceding the 2A peptide can improve cleavage efficiency.
Consider gene order: The order of genes flanking the 2A peptide can influence cleavage efficiency.
Avoid ad-hoc sequences: Inserting random sequences for cloning purposes can decrease cleavage efficiency.