Analytical Sciences

Effect of a Chloroacetyl Modification on the Suppression of Dissociation of a Fluorescent Molecule from Cells for Antigen-Specific Cell Staining

Ryosuke KANEKO,*1 Masumi KAWAMURA,*2 Akihiro KISHIMURA,*1,*2,*3,*4 Takeshi MORI,*1,*2,*3 and Yoshiki KATAYAMA*1,*2,*3,*4,*5,*6

*1 Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*2 Graduate School of System Life Science, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*3 Center for Future Chemistry, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*4 International Research Center for Molecular Systems, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*5 Center for Advanced Medical Innovation, Kyushu University, 3-1-1 Maidashi, Higashi, Fukuoka 812-8582, Japan
*6 Department of Biomedical Engineering, Chung Yuan Christian University, 200 Chung Pei Rd., Chung Li, 32023 ROC, Taiwan

We previously developed a hydrolase-based fluorescence amplification method for antigen-specific cell labelling, in which fluorescent substrates stained cells by a non-covalent hydrophobic interaction. To improve the substrates retention in cells, we examined the effect of a chloroacetyl group modification on the substrate retention. We found that the chloroacetyl group suppressed the dissociation of the substrate after forming a covalent bond with intracellular proteins. However, the slow reaction speed of the chloroacetyl group allowed dissociation for cells in the early stage of the staining reaction.

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