Carbohydrate monolayer strategy for electrochemical assay of cell surface carbohydrate.
The study of glycobiology has been seriously hampered due to lack of an ideal assay tool. This work proposes a robust carbohydrate monolayer platform to solve the problems of active site inaccessibility and lectin denaturation associated with protein arrays reported for detection of cell surface carbohydrates and develops a convenient method for monitoring cell surface carbohydrate sites of interest, with high sensitivity, acceptable rapidity, low cost, and excellent extensibility. It utilizes the competitive binding of solid-surface-confined and cell-surface-residing carbohydrates to quantum dot labeled carbohydrate recognition protein and subsequent voltammetric quantification of the metal signature. The mannan monolayer strategy exhibited sensitive response to K562 cells and possessed potential specificity due to the specific interaction between lectin and corresponding carbohydrate. By comparing the competitive binding of K562 cells with mannan in solutions, the average Con A binding capacity of a single K562 cell could be estimated to correspond to 6.9 pg or 2.3 x 10(10) mannose moieties. This strategy integrates the advantages of surface assembly, nanotechnology, bioconjugate techniques, and electrochemical detection and can be expanded for profiling cell surface carbohydrates and high-throughput multiple detection by simultaneously using more pairs of lectin and carbohydrate owing to the multiple coding capability of QDs, which provides an important protocol for the quantitative evaluation of cell surface carbohydrate sites.
Ding
Lin
L
Key Laboratory of Analytical Chemistry for Life Science, Ministry of Education of China, Department of Chemistry, Nanjing University, Nanjing 210093, PR China.
Cheng
Wei
W
Wang
Xiaojian
X
Ding
Shijia
S
Ju
Huangxian
H
D001665:Binding Sites; D001667:Binding, Competitive; D040761:Biomimetic Materials / Q000737:chemistry / Q000378:metabolism; D002241:Carbohydrates / Q000032:analysis*; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003208:Concanavalin A / Q000032:analysis / Q000378:metabolism; D004563:Electrochemistry / Q000379:methods*; D006801:Humans; D020014:K562 Cells; D008351:Mannans / Q000737:chemistry / Q000378:metabolism; D008358:Mannose / Q000032:analysis / Q000378:metabolism; D045663:Quantum Dots; D013379:Substrate Specificity
D002241:Carbohydrates; D008351:Mannans; D003208:Concanavalin A; D008358:Mannose