Lectin-Based Nanoprobes Functionalized with Enzyme for Highly Sensitive Electrochemical Monitoring of Dynamic Carbohydrate Expression on Living Cells

Abstract

A dual-functionalized nanoprobe was designed for highly sensitive and selective in situ evaluation of carbohydrates on living cells by integrating the specific carbohydrate recognition and enzymatic signal amplification of proteins on Au nanoparticles. A nanoscaffold of nanohorns functionalized with arginine-glycine-aspartic acid-serine tetrapeptide was also prepared on an electrode surface for cell capture and enhancing the electrical connectivity. Combined with the nanoprobe and peptide-nanohorns, a highly sensitive electrochemical strategy was developed for cytosensing, which showed a detection limit down to 15 cells, broad dynamic range, acceptable rapidity, and low cost. The proposed method was further used for monitoring of dynamic variation of carbohydrate expression on cancer cells in response to drugs, which obviated the destruction or labeling of cells and the covalent tagging of lectin and enzyme. The one-pot conjugation of three components was very convenient and could be extended for preparation of other lectin-functionalized nanoprobes. Further development of this technique would contribute considerably to meeting the challenges in comprehensive understanding of the glycomic codes.

A repeatable assembling and disassembling electrochemical aptamer cytosensor for ultrasensitive and highly selective detection of human liver cancer cells

“…The prepared AuNPs solution was stored at 4 °C for further use. 9 As shown in Scheme 1A, probe-AuNPs-HRP nanoprobes were firstly achieved by adding HRP and capture probe to an AuNPs solution [12]. Firstly, 50 μL of HRP (1 mg mL -1 ) were added to 2 mL AuNPs solution which was adjusted to pH 9.0.…”

“…Firstly, the HRP-AuNPs nanoprobe was prepared by incubation of AuNPs with HRP at pH 9 for 60 min under gentle stirring [12]. After the conjugation of AuNPs and HRP, we attached the thiolated DNA to HRP modified-AuNPs to form probeAuNPs-HRP nanoprobe with assistance of NaCl.…”

“…Recently, there have been some attempts for cancer cell detection using electrochemical detection methods, for example electrochemical impedance spectroscopy (EIS) [7][8][9][10], differential pulse voltammetry (DPV) [11][12][13][14], cyclic voltammetry (CV) [15], stripping voltammetry (SV) [16,17] and electrochemical luminescence (ECL) [18][19]. Compared with the typical methods, electrochemical methods do offer the advantages of acceptable sensitivity, operational simplicity, rapid response and low cost.…”

Sensitive biosensing strategy based on functional nanomaterials

“…By integrating the specific carbohydrate recognition and enzymatic signal amplification of proteins on Au nanoparticles, a dual-functionalized nanoprobe has been designed for highly sensitive and selective in situ evaluation of glycans on living cells [84]. This method uses a nano-scaffold of nanohorns functionalized with arginine-glycine-aspartic acid-serine tetrapeptide to capture cell and enhance electrical connectivity, which allows a detection limit down to 15 cells and monitoring of dynamic variation of carbohydrate expression on cancer cells in response to drugs.…”

Biofunctionalization of Nanomaterials

“…Recently, based on specific recognition between lectins and sugar epitopes, a concanavalin A-functionalized Au NP as nanoprobe was developed for the highly sensitive and selective in situ evaluation of carbohydrates on living cells by electrochemical tracing of the enzyme coimmobilized on the NPs [28]. Coupled with the efficient capture of cells on electrodes modified with arginine-glycine-aspartic acidserine tetrapeptide-functionalized SWNTs by the specific affinity interaction between the tetrapeptides and the integrins on the cell surface, the dynamic changes in four kinds of cell surface carbohydrates during drug treatment can be monitored by four horseradish peroxidase (HRP)-labeled lectins [29].…”