TY - JOUR
T1 - Preparation of affinity membranes using thermally induced phase separation for one-step purification of recombinant proteins
AU - Honjo, Takafumi
AU - Hoe, Kazuki
AU - Tabayashi, Shunsuke
AU - Tanaka, Tsutomu
AU - Shimada, Josui
AU - Goto, Masahiro
AU - Matsuyama, Hideto
AU - Maruyama, Tatsuo
N1 - Funding Information:
This work was supported financially by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan, by Grants for Research and Technology Development on Waste Management ( K2336 ) from the Ministry of Environment, Japan, and also by a Grant-in-Aid for Challenging Exploratory Research ( 23656491 ).
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2013/3/15
Y1 - 2013/3/15
N2 - We synthesized several surfactant-like ligands and prepared affinity membranes by introducing them into porous polymeric membranes using the thermally induced phase separation method. The ligands (nitrilotriacetate, iminodiacetate, and glutathione) were successfully displayed on the surfaces of cellulose diacetate membranes. Membranes functionalized with nitrilotriacetate and glutathione captured and released hexahistidine-tagged enhanced green fluorescent protein (His-tag GFP) and glutathione S-transferase (GST) selectively under appropriate conditions. The affinity membranes also enabled highly selective purification of target proteins (GFP and GST) from cell lysates. The protein-binding capacity was 15 μg/cm2 for His-tag GFP and 13 μg/cm2 for GST. The application-specific membranes described in this work will aid high-throughput screening and high-throughput analysis of recombinant proteins.
AB - We synthesized several surfactant-like ligands and prepared affinity membranes by introducing them into porous polymeric membranes using the thermally induced phase separation method. The ligands (nitrilotriacetate, iminodiacetate, and glutathione) were successfully displayed on the surfaces of cellulose diacetate membranes. Membranes functionalized with nitrilotriacetate and glutathione captured and released hexahistidine-tagged enhanced green fluorescent protein (His-tag GFP) and glutathione S-transferase (GST) selectively under appropriate conditions. The affinity membranes also enabled highly selective purification of target proteins (GFP and GST) from cell lysates. The protein-binding capacity was 15 μg/cm2 for His-tag GFP and 13 μg/cm2 for GST. The application-specific membranes described in this work will aid high-throughput screening and high-throughput analysis of recombinant proteins.
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U2 - 10.1016/j.ab.2012.11.027
DO - 10.1016/j.ab.2012.11.027
M3 - Article
C2 - 23274363
AN - SCOPUS:84873403315
SN - 0003-2697
VL - 434
SP - 269
EP - 274
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -