Preparation of Thrombin Aptasensor Based on the Metal-organic Framework Fe-MIL-88NH2†

doi:10.7503/cjcu20180563

Abstract

Abstract:

A sandwich-type aptasensor based on the metal-organic framework Fe-MIL-88NH₂ for the rapid detection of thrombin was developed. Amino-functionalized metal-organic framework(Fe-MOFs), Fe-MIL-88NH₂, was used as a label, which was covalently cross linked with amino-modified thrombin aptamer Ⅱ through glutaraldehyde. Gold nanoparticle modified covalent organic frameworks(Au NPs@COF-LZU-8) with high electronic conductivity was employed as the substrate to immobilize thiolated thrombin aptamer Ⅰ(TBAⅠ). After thrombin binds specifically with TBAⅠ, it combines with Fe-MIL-88NH₂ labeled thrombin aptamer Ⅱ. Then, iron ions(Fe³⁺) from Fe-MIL-88NH₂ were directly detected without acid dissolution and preconcentration. Under the optimized experimental conditions, the current response of Fe³⁺ in Fe-MIL-88NH₂ was directly proportional to the concentration of thrombin in the range of 0.5—200 ng/mL, with a linear correlation coefficient of 0.987 and a detection limit of 0.167 ng/mL. In this strategy, the detecting steps were greatly simplified. This article expands the application range of MOFs.

Key words: Thrombin, Fe-metal-organic framework, Fe-MIL-88NH₂, Aptasensor, Sandwich type

CLC Number:

O657.1

TrendMD:

LI Delei,GU Mengqiao,WANG Min,CHI Kuanneng,ZHANG Xi,DENG Yan,MA Yuchan,HU Rong,YANG Yunhui. Preparation of Thrombin Aptasensor Based on the Metal-organic Framework Fe-MIL-88N $H 2 †$ [J]. Chem. J. Chinese Universities, 2019, 40(3): 439.

Figures/Tables 13

Fig.1 3D structure(A) and planar structure(B) diagrams of Fe-MIL-88NH2

Fig.2 Stepwise procedure of the aptasensor

Fig.3 TEM images of COF-LZU-8(A) and Au NPs@COF-LZU-8(B)

Fig.4 XRD pattern of COF-LZU-8

Fig.5 SEM images of Fe-MIL-88NH2

Fig.6 XRD patterns of standard(A) and synthesized(B) Fe-MIL-88NH2

Fig.7 DPV response curves of interfaces at diffe-rent modified electrode in PBS buffer solutiona. TB/TBAⅠ/Au NPs@COF-LZU-8/Chit/GCE; b. TBAⅡ-Fe-MIL-88NH2/TBAⅠ/Au NPs@COF-LZU-8/Chit/GCE; c. TBAⅡ-Fe-MIL-88NH2/TB/TBAⅠ/Au NPs@COF-LZU-8/Chit/GCE.

Fig.8 Electrochemical impedance spectroscopy(EIS) of interfaces at different modified electrode in 5 mmol/L K3Fe(CN)6/K4Fe(CN)6a. Bare GCE; b. Au NPs@COF-LZU-8/Chit/GCE; c. TBAⅠ/Au NPs@COF-LZU-8/Chit/GCE; d. MCH/TBAⅠ/Au NPs@COF-LZU-8/Chit/GCE; e. TB/MCH/TBAⅠ/Au NPs@COF-LZU-8/Chit/GCE; f. TBAⅡ-Fe-MIL-88NH2/TB/MCH/TBAⅠ/Au NPs@COF-LZU-8/Chit/GCE.

Fig.9 Effects of buffer type(A), pH values(B), TBAⅠconcentration(C) and the incubation time(D) on sensing signals of the aptasensor between TB and TBAⅡ-Fe-MIL-88NH2 bioconjugates

Fig.10 Calibration curve of aptasensor in PBS solution(A) and DPV response of aptasensor to different concentrations of TB(B)(B) c(TB)/(ng·mL-1), a.—i.: 0, 0.5, 1, 5, 10, 25, 50, 100, 200.

Table 1 Comparison of different detecting methods for TB

No.	Method	Linear range/ (nmol·L^-1)	Detection limit/ (nmol·L^-1)	Ref.
1	Electrochemical aptasensor	0.099—55	0.099	[26]
2	Electrochemical aptasensor	0.298—49.5	0.0099	[27]
3	Label-free electrochemical aptasensor	0.12—46	0.4	[28]
4	Exonuclease Ⅲ-aided electrochemical detection(DPV)	0.005—1.0	0.005	[29]
	SWV	0.010—10.0	0.0012	[30]
5	Electrochemical aptasensor	0.0135—5.4	0.0045	This work

Fig.11 Selectivity of the aptasensor

Table 2 Recovery of aptasensor

Sample	Added/ (ng·mL^-1)	Found/ (ng·mL^-1)	RSD(%)	Recovery (%)	Sample	Added/ (ng·mL^-1)	Found/ (ng·mL^-1)	RSD(%)	Recovery (%)
1	0	0.31	15.3	3		10.00	9.33		90.7
	0	0.24			4	50.00	51.42	3.02	102.3
	0	0.24				50.00	53.34		106.2
2	5.00	5.08	2.20	96.6		50.00	50.95		101.4
	5.00	5.29		100.6	5	100	97.6	1.90	97.3
	5.00	5.11		97.0		100	101.2		100.9
3	10.00	10.04	4.05	97.8		100	100.4		100.1
	10.00	9.99		97.3

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