title: Interaction evaluation of bacteria and protoplasts with single-stranded deoxyribonucleic acid library based on capillary electrophoresis.
abstract: For whole-cell aptamers selection, cells surface situation has great impact on single-stranded (ssDNA) binding and aptamers selection. In this work, both Lactobacillus acidophilus and Escherichia coli as well as their protoplasts were as cells targets, their interaction with ssDNA library were evaluated based on capillary zone electrophoresis (CZE) and affinity capillary electrophoresis (ACE) with UV and LIF detection. Our results demonstrated that protoplasts without cells wall had apparently stronger interaction with ssDNA library than bacteria, the protoplasts-ssDNA complex could be observed clearly with CZE-LIF. Furthermore, E. coli pretreated by four organic solvents (methanol, ethanol, formaldehyde and glutaraldehyde) showed binding difference with ssDNA library, which could be identified with ACE-UV. Binding constants indicated the interaction of E. coli with ssDNA library were in the order of E. coli protoplasts>methanol (ethanol) treated E. coli>formaldehyde (glutaraldehyde) treated E. coli≈E. coli. Above results suggest that cells surface situation determines their binding affinity with ssDNA, which should be considered in whole-cell aptamers selection and aptamers further application. Capillary electrophoresis is a preferable technique for interaction evaluation of composite targets binding with ssDNA library.
authors.affiliation: School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing 100081, China; State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Chaoyang
authors.initials: C
authors.lastname: Meng
authors.affiliation: Beijing Centre for Physical and Chemical Analysis, Beijing 100089, China.
authors.forename: Xinying
authors.initials: X
authors.lastname: Zhao
authors.affiliation: School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing 100081, China. Electronic address: qufengqu@bit.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Qu
authors.affiliation: School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing 100081, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Mei
authors.affiliation: School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Beijing 100081, China.
authors.forename: Li
authors.initials: L
authors.lastname: Gu
mesh_terms: D004277:DNA, Single-Stranded / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry* / Q000166:cytology; D005557:Formaldehyde / Q000737:chemistry; D015723:Gene Library; D005976:Glutaral / Q000737:chemistry; D007779:Lactobacillus acidophilus / Q000737:chemistry* / Q000166:cytology; D011523:Protoplasts / Q000737:chemistry; D052156:SELEX Aptamer Technique; D012997:Solvents / Q000737:chemistry
chemical_list: D004277:DNA, Single-Stranded; D012997:Solvents; D005557:Formaldehyde; D005976:Glutaral
keywords: Aptamers selection; Bacteria; Capillary electrophoresis; Interaction; Protoplasts; ssDNA