Products/Services Used | Details | Operation |
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Monoclonal Antibody Services> | HA tag (YPYDVPDYA)-fused scFv constructs (s3Flag-scFv-HA) were synthesised and codon-optimised for expression in mice using Genscript. | Get A Quote |
Targeting cytoplasmic protein-protein interactions with antibodies remains technically challenging, since antibodies expressed in the cytosol frequently form insoluble aggregates. Existing engineering methods are based on the notion that the estimated net charge at pH 7.4 affects stability; as such, they are unable to overcome this problem. Herein, we report a versatile method for engineering an ultra-stable cytoplasmic antibody (STAND), with a strong estimated net negative charge at pH 6.6, by fusing peptide tags with a highly negative charge and a low isoelectric point. Without the need for complicated amino acid substitutions, we convert aggregation-prone antibodies to STANDs that are useful for inhibiting i... More