Synthesis · Laccase-inspired bi-amino acid MOFs with high substrate affinity: Catalytic deposition induced “signal-down” electrochemical response towards PD-L1

4 structured synthesis routes

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

Complete recipeSource: Main

Route 1: Other

4 · 2.2

Metal precursorsCu-F/Hs solution (2 mg/mL)

Solventsaqueous buffer/solution not further specified

AdditivesPD-L1 antibody, EDC, NHS

Atmospherenot specified

Temperatureroom temperature

Time2.5

Work-upCentrifuged at 8000 rpm for 10 min and washed.

Scalability contextSolution-phase bioconjugation recipe at millilitre scale.

Show 4 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	PD-L1 Ab solution	1 mL · 100 ng/mL	4 · 2.2
Additive	EDC solution	500 uL · 24 mg/mL	4 · 2.2
Additive	NHS solution	500 uL · 16 mg/mL	4 · 2.2
Other	Cu-F/Hs solution	2 mL · 2 mg/mL	4 · 2.2

Complete recipeSource: Main

Route 2: Solvothermal

3 · 2.2

Metal precursors2 mL Cu(NO3)2 solution (0.24 M)

Linker precursors2 mL His solution (0.12 M) and 2 mL Phe solution (0.12 M)

Solvents8 mL DMF; aqueous/deionised water used in workup

Atmospherenot specified

Temperature100

Time1

Work-upPrecipitate washed with DMF and deionised water, dried.

ActivationStored at 4 degC for later use; no activation reported.

Scalability contextSmall-batch autoclave recipe; 14 mL total precursor/solvent volume reported.

Show 4 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Cu(NO3)2 solution	2 mL · 0.24 M	3 · 2.2
Linker	His solution	2 mL · 0.12 M	3 · 2.2
Linker	Phe solution	2 mL · 0.12 M	3 · 2.2
Solvent	DMF	8 mL	3 · 2.2

Complete recipeSource: SI

Route 3: Solvothermal

2 · 1.1 Synthesis of Cu-Fs

Metal precursors2 mL Cu(NO3)2 solution (0.24 M)

Linker precursors4 mL Phe solution (0.12 M)

Solvents8 mL DMF; DMF and deionised water workup

Atmospherenot specified

Temperature100

Time1

Work-upPrecipitate washed with DMF and deionised water, dried.

ActivationStored at 4 degC for later use; no activation reported.

Scalability contextControl recipe uses the same total ligand volume as Cu-F/Hs but only Phe.

Show 3 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	Cu(NO3)2 solution	2 mL · 0.24 M	2 · 1.1 Synthesis of Cu-Fs
Linker	Phe solution	4 mL · 0.12 M	2 · 1.1 Synthesis of Cu-Fs
Solvent	DMF	8 mL	2 · 1.1 Synthesis of Cu-Fs

Complete recipeSource: Main

Route 4: Electrochemical

4 · 2.3-2.4 · Scheme 1B

Metal precursorsgold working electrode; Ab-Cu-F/Hs contains Cu-F/Hs

Linker precursorsPD-L1 aptamer and PD-L1 antibody-bearing Cu-F/Hs

Solvents0.5 M H2SO4 for electrode cycling; aqueous assay solutions

AdditivesTCEP, MCH, PD-L1, DA, ODA

Atmospherenitrogen drying after gold-electrode cleaning; otherwise not specified

Temperatureroom temperature

Timeabout 4

Substrate orientationgold electrode surface

Oxidant / reductantDA oxidatively polymerised/deposited during Cu-F/Hs-catalysed reaction with ODA

Work-upGold electrode polished, sonicated, piranha-cleaned, rinsed with deionised water and dried with nitrogen before modification.

ActivationGold electrode cycled 20 times from 0 to 1.6 V in 0.5 M H2SO4.

Scalability contextSingle-electrode biosensor assembly; volumes 50-100 uL for assay additions.

Show 7 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	PD-L1 Apt	100 uL applied after TCEP treatment · 2 uM stock mixed 1:1 with TCEP; final selected Apt concentration 1 uM	4 · 2.3-2.4 · Scheme 1B
Additive	TCEP	1:1 with PD-L1 Apt · 5 mM	4 · 2.3-2.4 · Scheme 1B
Additive	MCH	100 uL · 10 mM	4 · 2.3-2.4 · Scheme 1B
Other	PD-L1	50 uL	4 · 2.3-2.4 · Scheme 1B
Other	Ab-Cu-F/Hs	100 uL	4 · 2.3-2.4 · Scheme 1B
Other	DA	50 uL · 10 uM	4 · 2.3-2.4 · Scheme 1B
Additive	ODA	50 uL · 100 uM	4 · 2.3-2.4 · Scheme 1B