Preparation of affinity membranes using thermally induced phase separation for one-step purification of recombinant proteins

Takafumi Honjo, Kazuki Hoe, Shunsuke Tabayashi, Tsutomu Tanaka, Josui Shimada, Masahiro Goto, Hideto Matsuyama, Tatsuo Maruyama

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)269-274
Number of pages6
JournalAnalytical Biochemistry
Volume434
Issue number2
DOIs
Publication statusPublished - Mar 15 2013

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Recombinant Proteins
His-His-His-His-His-His
Phase separation
Purification
Membranes
Glutathione Transferase
Glutathione
Throughput
Ligands
Polymeric membranes
Surface-Active Agents
Protein S
Screening
Protein Binding
Proteins
enhanced green fluorescent protein

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Preparation of affinity membranes using thermally induced phase separation for one-step purification of recombinant proteins. / Honjo, Takafumi; Hoe, Kazuki; Tabayashi, Shunsuke; Tanaka, Tsutomu; Shimada, Josui; Goto, Masahiro; Matsuyama, Hideto; Maruyama, Tatsuo.

In: Analytical Biochemistry, Vol. 434, No. 2, 15.03.2013, p. 269-274.

Research output: Contribution to journalArticle

Honjo, Takafumi ; Hoe, Kazuki ; Tabayashi, Shunsuke ; Tanaka, Tsutomu ; Shimada, Josui ; Goto, Masahiro ; Matsuyama, Hideto ; Maruyama, Tatsuo. / Preparation of affinity membranes using thermally induced phase separation for one-step purification of recombinant proteins. In: Analytical Biochemistry. 2013 ; Vol. 434, No. 2. pp. 269-274.
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