TY - JOUR
T1 - Column-free purification of an artificial protein nanocage, TIP60
AU - Nasu, Erika
AU - Kawakami, Norifumi
AU - Ohara, Naoya
AU - Hayashi, Keiichi
AU - Miyamoto, Kenji
N1 - Funding Information:
We thank Prof. Y. Watanabe and Prof. O. Shoji of Nagoya University for the DLS measurements. This work is financially supported by JSPS KAKENHI , Grant Number JP18K05324 to N. K., and by JST SPRING, Grant Number JPMJSP2123 to E.N.
Publisher Copyright:
© 2023 Elsevier Inc.
PY - 2023/5
Y1 - 2023/5
N2 - Protein nanocages, which have inner cavities and surface pores, are attractive materials for various applications, such as in catalysts and medicine. Recently, we produced an artificial protein nanocage, TIP60, and demonstrated its potential as a stimuli-responsive nanocarrier. In the present study, we report a simple purification method for TIP60 that can replace time-consuming and costly affinity chromatography purification. TIP60, which has an anionic surface charge, aggregated at mildly acidic pH and redissolved at neutral pH, maintaining its cage structure. This pH-responsive reversible precipitation allowed us to purify TIP60 from soluble fractions of the E. coli cell lysate by controlling the pH. Compared with conventional Ni-NTA column purification, the pH-responsive precipitation method provided purified TIP60 with similar purity (∼80%) and higher yield. This precipitation purification method should facilitate the large-scale investigation and practical use of TIP60 nanocages.
AB - Protein nanocages, which have inner cavities and surface pores, are attractive materials for various applications, such as in catalysts and medicine. Recently, we produced an artificial protein nanocage, TIP60, and demonstrated its potential as a stimuli-responsive nanocarrier. In the present study, we report a simple purification method for TIP60 that can replace time-consuming and costly affinity chromatography purification. TIP60, which has an anionic surface charge, aggregated at mildly acidic pH and redissolved at neutral pH, maintaining its cage structure. This pH-responsive reversible precipitation allowed us to purify TIP60 from soluble fractions of the E. coli cell lysate by controlling the pH. Compared with conventional Ni-NTA column purification, the pH-responsive precipitation method provided purified TIP60 with similar purity (∼80%) and higher yield. This precipitation purification method should facilitate the large-scale investigation and practical use of TIP60 nanocages.
KW - pH-responsive reversible precipitation
KW - Protein nanocage
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U2 - 10.1016/j.pep.2023.106232
DO - 10.1016/j.pep.2023.106232
M3 - Article
C2 - 36642237
AN - SCOPUS:85146871314
SN - 1046-5928
VL - 205
JO - Protein Expression and Purification
JF - Protein Expression and Purification
M1 - 106232
ER -