Versatile aptasensor for electrochemical quantification of cell surface glycan and naked-eye tracking glycolytic inhibition in living cells.
Quantifying the glycan expression status on cell surfaces is of vital importance for insight into the glycan function in biological processes and related diseases. Here we developed a versatile aptasensor for electrochemical quantification of cell surface glycan by taking advantage of the cell-specific aptamer, and the lectin-functionalized gold nanoparticles acting as both a glycan recognition unit and a signal amplification probe. To construct the aptasensor, amine-functionalized mucin 1 protein (MUC1) aptamer was first covalently conjugated to carboxylated-magnetic beads (MBs) using the succinimide coupling (EDC-NHS) method. On the basis of the specific recognition between aptamer and MUC1 protein that overexpressed on the surface of MCF-7 cells, the aptamer conjugated MBs showed a predominant capability for cell capture with high selectivity. Moreover, a lectin-based nanoprobe was designed by noncovalent assembly of concanavalin A (ConA) on gold nanoparticles (AuNPs). This nanoprobe incorporated the abilities of both the specific carbohydrate recognition and the signal amplification based on the gold-promoted reduction of silver ions. By coupling with electrochemical stripping analysis, the proposed sandwich-type cytosensor showed an excellent analytical performance for the ultrasensitive detection of MCF-7 cells and quantification of cell surface glycan. More importantly, taking advantage of Con A-gold nanoprobe catalyzed silver enhancement, the proposed method was further used for naked-eye tracking glycolytic inhibition in living cells. This aptasensor holds great promise as a new point-of-care diagnostic tool for analyzing glycan expression on living cells and further helps cancer diagnosis and treatment.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China; Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Jing-Jing
JJ
Zhang
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China; School of Pharmacy, Nanjing University of Chinese Medicine, 210023, China.
Fang-Fang
FF
Cheng
Department of Chemistry, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241, China. Electronic address: ttzheng@chem.ecnu.edu.cn.
Ting-Ting
TT
Zheng
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China. Electronic address: jjzhu@nju.edu.cn.
Jun-Jie
JJ
Zhu
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques; D003208:Concanavalin A / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D061986:MCF-7 Cells; D053768:Metal Nanoparticles; D018396:Mucin-1 / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification*; D013388:Succinimides / Q000737:chemistry
D052157:Aptamers, Nucleotide; C060192:MUC1 protein, human; D018396:Mucin-1; D011134:Polysaccharides; D013388:Succinimides; C083511:concanavalin A-gold; C032620:succinimide; D003208:Concanavalin A
Aptamer; Electrochemical sensors; Glycan analysis; Gold nanoprobse; Naked-eye detection