Stability towards alkaline conditions can be engineered into a protein ligand
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Published on: February 11, 2019
S Gülich1, M Linhult, P Nygren
1Department of Biotechnology, Royal Institute of Technology, SE-100 44, Stockholm, Sweden.
Abstract:
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 in 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.
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