| Autor: |
Daiki Sugihara, Fuka Ono, Motoki Sugino, Hiromi Suzuki, Noriko Endo, Atsuhiro Shimada, Akio Ebihara |
| Předmět: |
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| Zdroj: |
Bioscience, Biotechnology & Biochemistry; Sep2023, Vol. 87 Issue 9, p1029-1035, 7p |
| Abstrakt: |
Triple-FLAG (3 × FLAG)-tagged proteins can be affinity purified through binding to an anti-FLAG antibody and competitive elution with excess free 3 × FLAG peptide. To expand the availability of the 3 × FLAG purification system, we produced a recombinant His-tagged 3 × FLAG peptide in Brevibacillus choshinensis. The screening of connecting linkers between His-tag and the 3 × FLAG peptide, culture containers, and culture media showed that the His-tagged 3 × FLAG peptide with an LA linker was most expressed in 2SY medium using a baffled shake flask. The peptide was affinity-purified to give a yield of about 25 mg/L of culture. The peptide was effective for eluting 3 × FLAG-tagged α-amylase from anti-FLAG magnetic beads. Finally, the peptide remaining in the amylase fraction was removed by His-tag affinity purification. These results show that the recombinant His-tagged 3 × FLAG peptide can function as an easy-to-remove affinity peptide in the 3 × FLAG purification system. The recombinant His-tagged 3 × FLAG peptide produced using the Brevibacillus expression system can expand the utility of the 3 × FLAG purification system. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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