Stability towards alkaline conditions can be engineered into a protein ligand

• One of the problems with a proteinaceous affinity ligand is their sensitivity to alkaline conditions. Here, we show that a simple and straightforward strategy consisting of replacing all asparagine residues with other amino acids can dramatically improve the chemical stability of a protein towards alkaline conditions. As a model, a Streptococcal albumin-binding domain (ABD) was used. The engineered variant showed higher stability towards 0.5 M NaOH, as well as higher thermal stability compared to its native counterpart. This protein engineering approach could potentially also be used for other protein ligands to eliminate the sensitivity to alkaline cleaning-in-place (CIP) conditions.

Nyckelord

albumin-binding domain

cleaning-in-place

deamidation

human serum albumin

protein engineering

serum albumin

deamidation

residues

asparaginyl

peptides

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)