Synthesis · Sensitive detection of carcinoembryonic antigen (CEA) by a sandwich-type electrochemical immunosensor using MOF-Ce@HA/Ag-HRP-Ab2 as a nanoprobe

5 structured synthesis routes

Completeness describes how fully the route could be reconstructed from the main article and supporting information.

Partial recipeSource: Main

Route 1: Other

3 · 2.6

Metal precursorsCe-MoF@HA/Ag

SolventsPBS washing solution; EDC/NHS aqueous solutions implied but solvent not specified

AdditivesEDC; NHS; Ab2; HRP; BSA

Atmospherenot specified

Temperature4

TimeEDC/NHS storage 6; HRP incubation 12; BSA 2; Ce-MoF@HA/Ag stirring 12

Work-upCentrifuged at 4 C and washed three times with PBS solution.

ActivationEDC/NHS activation before Ab2 and HRP addition

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Additive	EDC	0.5 ml · 0.1 mol l-1	3 · 2.6
Additive	NHS	mixed with EDC, v1:v2 = 1:4 · 0.1 mol l-1	3 · 2.6
Other	Ab2	0.5 ml	3 · 2.6
Other	HRP	0.5 ml · 4 mg ml-1	3 · 2.6
Additive	BSA	200 ul · 1 wt%	3 · 2.6
Other	Ce-MoF@HA/Ag	0.5 ml · 4 mg ml-1	3 · 2.6

Partial recipeSource: Main

Route 2: Other

3 · 2.5

Metal precursorsCe-MoF@HA; AgNO3

Solventsdeionized water

Atmospherenot specified

Temperatureice-water bath; standing at 4

TimeNaBH4 stirring 0.267; standing 12

Oxidant / reductantAgNO3 reduced by NaBH4

Work-upProduct washed three times with deionized water.

Show 4 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	Ce-MoF@HA	25 mg	3 · 2.5
Solvent	deionized water	15 ml	3 · 2.5
Metal Source	AgNO3	7 ml · 0.001 mol l-1	3 · 2.5
Reductant	NaBH4	20 ml · 0.01 mol l-1	3 · 2.5

Partial recipeSource: Main

Route 3: Other

3 · 2.4

Metal precursorsCe-MoF from route_ce_mof_water_bath

Linker precursorshyaluronic acid (HA)

SolventsDMF-water mixture: 23 ml DMF and 2 ml deionized water

Atmospherenot specified

Temperatureroom temperature for stirring; drying 70

Timesonication 0.25; stirring overnight; drying 12

Work-upPurified product washed three times with DMF and acetone.

Activationdried at 70 C for 12 h

Show 4 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	Ce-MoF	25 mg	3 · 2.4
Additive	hyaluronic acid (HA)	25 mg	3 · 2.4
Solvent	DMF	23 ml	3 · 2.4
Solvent	deionized water	2 ml	3 · 2.4

Partial recipeSource: Main

Route 4: Other

3 · 2.3

Metal precursors0.217 g Ce(NO3)3.6H2O (5 mM) in 5 ml deionized water

Linker precursors0.105 g H3BTC (5 mM) in 5 ml ethanol-water (1:1)

Solventsdeionized water; ethanol-water mixed solution (v1:v2 = 1:1)

Atmospherenot specified

Temperature60; drying 70

Time1; drying 12

Work-upCe nitrate solution added dropwise to H3BTC solution; vigorous stirring; milky white precipitate washed three times with ethanol and deionized water.

Activationdried at 70 C for 12 h and stored for use

Show 4 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Ce(NO3)3.6H2O	0.217 g · 5 mM	3 · 2.3
Linker	H3BTC	0.105 g · 5 mM	3 · 2.3
Solvent	deionized water	5 ml	3 · 2.3
Solvent	ethanol-water mixed solution	5 ml · v1:v2 = 1:1	3 · 2.3

Partial recipeSource: Main

Route 5: Drop Cast

3-4 · 2.7 · Figure 1

Metal precursorsAu nanoparticles; MoF-Ce@HA/Ag-HRP-Ab2

Solventsabsolute ethanol; deionized water; 0.1 mol PBS pH 7.0 rinsing solution

AdditivesAb1; BSA; CEA antigen

Atmosphereair / room temperature where stated

Temperature4 for Ab1 and BSA incubation; room temperature for CEA incubation; storage 4

TimeAb1 10; BSA 3; CEA 0.667; nanoprobe 0.333

Substrate orientation4 mm glassy carbon electrode

Work-upGCE polished with 0.5 um and 50 nm Al2O3, washed three times with ethanol and deionized water; electrode rinsed with 0.1 mol PBS pH 7.0 after each step.

ActivationAu NPs used as platform for Ab1 immobilisation; BSA blocking before CEA and nanoprobe incubation.

Show 5 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	Au NPs	8 ul	3-4 · 2.7 · Figure 1
Other	Ab1	10 ul · 22 ng ml-1	3-4 · 2.7 · Figure 1
Additive	BSA	1%	3-4 · 2.7 · Figure 1
Other	CEA antigen	different concentrations	3-4 · 2.7 · Figure 1
Other	MoF-Ce@HA/Ag-HRP-Ab2 nanoprobe	2 mg ml-1	3-4 · 2.7 · Figure 1