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Protein cell‐surface display through in situ enzymatic modification of proteins with a poly(Ethylene glycol)‐lipid
Author(s) -
Tomita Urara,
Yamaguchi Satoshi,
Maeda Yasukazu,
Chujo Kazuki,
Minamihata Kosuke,
Nagamune Teruyuki
Publication year - 2013
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.24933
Subject(s) - ethylene glycol , amphiphile , chemistry , peg ratio , biochemistry , cell , reagent , cancer cell , biophysics , microbiology and biotechnology , biology , organic chemistry , polymer , finance , cancer , economics , copolymer , genetics
Cell‐surface display of functional proteins is a powerful and useful tool for regulating and reinforcing cellular functions. Direct incorporation of site‐specifically lipidated proteins from the extracellular medium is more rapid, easily controllable and reliable in displaying active proteins than expression through gene transfer. However, undesirable amphiphilic reagents such as organic co‐solvents and detergents were required for suppressing aggregation of ordinary lipidated proteins in solution. We report here sortase A‐catalyzed modification of proteins with a poly(ethylene glycol)(PEG)‐lipid in situ on the surface of living cells. Proteins fused with a recognition tag were site‐specifically ligated with the PEG‐lipid which was preliminary incorporated into cell membranes. Accordingly, target proteins were successfully displayed on living cells without aggregation under an amphiphilic reagent‐free condition. Furthermore, to demonstrate the availability of the present method, Fc domains of immunoglobulin G were displayed on cancer cells, and the phagocytosis of cancer cells with dendritic cells were enhanced through the Fc–Fc receptor interaction. Thus, the present facile chemoenzymatic method for protein display can be utilized for modulating cell–cell interactions in cell and tissue engineering fields. Biotechnol. Bioeng. 2013;110: 2785–2789. © 2013 Wiley Periodicals, Inc.