Stability towards alkaline conditions can be engineered into a protein ligand

Switching an anti-IgG binding site between archaeal extremophilic proteins results in Affitins with enhanced pH stability

“…This has been well described for several proteins (Gulich et al, 2000(Gulich et al, , 2002Linhult et al, 2004). However, our previous attempts to improve the alkaline stability of Affitins by this method were unsuccessful (unpublished results).…”

Evaluation of commercial chromatographic adsorbents for the direct capture of polyclonal rabbit antibodies from clarified antiserum

“…In parallel with the development of new synthetic alternatives to Protein A, others have sought to improve Protein A adsorbents, largely through applying recombinant DNA techniques to: improve the ligand's tolerance to cleaning with sodium hydroxide [15,16]; lower the binding affinity to enable use of milder elution conditions [17]; and allowing Table 1 Description of adsorbents employed in this study for chemoselective oriented immobilisation to the base matrix through stable linkage chemistries [18].…”

Purification Systems Based on Bacterial Surface Proteins

“…The alkaline tolerance of the immunoglobulin-binding domains of SPG is not as good as for SPA, however resistance of proteins to environmental challenges can be substantially improved by protein engineering. In a strategy that was first proven effective for the albumin-binding domain of SPG, all four asparagine residues susceptible to base catalyzed deamidation were replaced with other residues (Gulich et al 2000a). This approach resulted in a molecule that could withstand repeated cycles of cleaning with high concentrations of sodium hydroxide, without any considerable loss in performance when linked to a chromatographic resin.…”

“…This approach resulted in a molecule that could withstand repeated cycles of cleaning with high concentrations of sodium hydroxide, without any considerable loss in performance when linked to a chromatographic resin. An increased thermal stability was also achieved without significant alteration in structure or function (Gulich et al 2000a). To improve the performance of this ligand even further, different linker regions and multimeric formats have been evaluated for tolerance to extended time periods of alkali exposure (Linhult et al 2003).…”