Degradation of C-terminal tag sequences on domain antibodies purified from E. coli supernatant.

ABSTRACT: Expression of recombinant proteins often takes advantage of peptide tags expressed in fusion to allow easy detection and purification of the expressed proteins. However, as the fusion peptides most often are flexible appendages at the N- or C-terminal, proteolytic cleavage may result in removal of the tag sequence. Here, we evaluated the functionality and stability of 14 different combinations of commonly used tags for purification and detection of recombinant antibody fragments. The tag sequences were inserted in fusion with the c-terminal end of a domain antibody based on the HEL4 scaffold in a phagemid vector. This particular antibody fragment was able to refold on the membrane after blotting, allowing us to detect c-terminal tag breakdown by use of protein A in combination with detecti

SUBMITTER: Lykkemark S 

PROVIDER: S-EPMC4622476 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature