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A chemically modified glass surface that facilitates transglutaminase-mediated protein immobilization |
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| Authors: | Yusuke Tanaka Satoshi Doi Noriho Kamiya Noriyuki Kawata Shinji Kamiya Kenichi Nakama Masahiro Goto |
| Affiliation: | (1) Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Fukuoka 819-0395, Japan;(2) Center for Future Chemistry, Kyushu University, 744 Motooka, Fukuoka 819-0395, Japan;(3) New Products & Buisiness Development Department, Nippon Sheet Glass Co., Ltd., 5-4 Tokodai, Tsukuba Ibaraki, 300-2635, Japan |
| Abstract: | An amino-modified glass surface for enzymatic protein immobilization by microbial transglutaminase (MTG) was developed. Diamine substrates with secondary amino groups in the linker moiety, like triethylenetetramine (TETA), exhibited at most a 2-fold higher reactivity in the MTG-catalyzed reaction compared to those with the alkyl linker. A 96-well glass plate was subsequently modified with selected diamine substrates. Validation of the modified surface by enzymatic immobilization of enhanced green fluorescent protein tagged with a glutamine donor-substrate peptide (LLQG) of MTG revealed that the protein loading onto the TETA-modified glass surface was approximately 15-fold higher than that on the unmodified one. |
| Keywords: | 96-Well glass plate Peptide tag Protein immobilization Site-specific protein modification Transglutaminase |
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