Stability towards alkaline conditions can be engineered into a protein ligand

作者：Gulich S; Linhult M; Nygren PA; Uhlen M; Hober S^*

来源：Journal of Biotechnology, 2000, 80(2): 169-178.

DOI：10.1016/S0168-1656(00)00259-5

• albumin-binding domain
• cleaning-in-place
• deamidation
• human serum albumin
• protein engineering

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.

• 出版日期2000-6-23