Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins

• The widely used His6 tag displays affinity to bare silica matrices. His6-tagged proteins bind specifically particles. Bare matrices are suitable for the purification of proteins. Double tagging with and Car9 improves binding strength. is advantageous single-step immobilization. “Green” protein purification/immobilization processes based on low-cost, earth-abundant, eco-friendly highly desirable. Unmodified fit well these demands. Since histidine-rich silica-binding peptides frequently isolated in biopanning experiments, this work aimed at assessing viability using as an alternative matrix His-tagged Adsorption desorption studies a purified EGFP shown that particles different size porosity occurred under conditions tested, elution could be accomplished eluants containing L-arginine/L-lysine. Non-tagged did not Small-scale batch schemes gel Davisil grade 643 or 646 Tris-buffered saline eluant 0.5 M L-arginine (pH 8.5) allowed purifying His6-EGFP from Escherichia coli lysates purity up 96% recovery yield ?70% after just one step. tagged peptide was recovered comparable yield. Other also similar levels. scale scheme extendable. These results demonstrate unmodified can effectively purify double His6-EGFP-Car9 only 30–55%, combination tags revealed immobilization purposes.