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Purification of MBP‐ and Strep ‐tag II‐tagged proteins
Author(s) -
Carlsson Marianne,
Heijbel Anna,
Karlsson Anneli,
Lilja Lydia
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.577.6
Subject(s) - chemistry , chromatography , sepharose , affinity chromatography , maltose binding protein , dextrin , yield (engineering) , peptide , biochemistry , recombinant dna , enzyme , materials science , starch , metallurgy , fusion protein , gene
Recombinant tagged proteins can be purified to high purity in one step using affinity chromatography. Two chromatography media are presented, StrepTactin™ Sepharose™ High Performance (HP) which has high affinity for the small 8 amino acid peptide, Strep ‐tag II, and Dextrin Sepharose HP, that binds to Maltose Binding Protein‐tag (MBP). The specificity of the two media towards their respective tag gives high purity of the target proteins. In this study, simple purifications using MBPTrap™ HP (prepacked with Dextrin Sepharose HP and StrepTrap™ HP, (prepacked with StrepTactin Sepharose HP) are presented. Reproducibility during repeated usage of StrepTrap HP was tested by running 6 repeated purifications of a Strep ‐tag II‐tagged protein expressed in E. coli on the same StrepTrap HP column including regeneration with 0.5 M NaOH between each run. No tendencies in reduction of final purity and yield of the target protein were observed. Equivalent results are obtained when scaling up showing the simplicity to purify larger quantities of pure target protein.