Synthesis · Construction of nanozyme based with mixed valence manganese oxide loaded on defective metal-organic frameworks for sensitive detection of biomarker procalcitonin

11 structured synthesis routes

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

Complete recipeSource: SI

Route 1: Other

SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP

Metal precursorsmagnetic balls (MBs), 5.00 mg

SolventsPBS buffer, 2.0 mL; PBST wash buffer; BSA storage/blocking solutions

AdditivesEDC 100 uL of 10.0 mg mL-1; NHS 50.0 uL of 10.0 mg mL-1; Ab1 20.0 uL of 5.00 mg mL-1; BSA 3.0% and 0.10%

Atmospherenot stated

TemperatureRT for activation/incubation; 4 C storage

Time0.5 h EDC/NHS activation; 1 h Ab1 incubation; 0.5 h BSA blocking

Work-upCarboxyl-activated MBs washed twice with PBS using a magnet; after Ab1 incubation, washed three times with PBST; after BSA blocking, washed three times with PBST.

ActivationEDC/NHS activation of carboxylated MBs for 30 min; BSA blocking after antibody conjugation.

Scalability contextSmall-batch bioconjugation in PBS.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	magnetic balls (MBs)	5.00 mg	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Other	PBS buffer	2.0 mL	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Additive	EDC	100 uL · 10.0 mg mL-1	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Additive	NHS	50.0 uL · 10.0 mg mL-1	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Other	PCT capture antibody (Ab1)	20.0 uL · 5.00 mg mL-1	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Other	BSA solution	2.0 mL blocking; 1.0 mL storage · 3.0%; 0.10%	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP

Complete recipeSource: SI

Route 2: Other

SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP

Metal precursorsMdP nanozyme, 10.0 mg

SolventsPBS buffer, 2.0 mL; PBST wash buffer; BSA storage/blocking solutions

AdditivesEDC 100 uL of 10.0 mg mL-1; NHS 50.0 uL of 10.0 mg mL-1; Ab2 40.0 uL of 5.00 mg mL-1; BSA 3.0% and 0.10%

Atmospherenot stated

TemperatureRT for EDC/NHS activation and antibody incubation; 4 C storage

Time0.5 h EDC/NHS activation; 1 h Ab2 incubation; 0.5 h BSA blocking

Work-upAfter EDC/NHS activation, wash twice with PBS by centrifugation (8000 rpm, 10 min); after Ab2 incubation, remove unbound Ab2 by washing three times with PBST (8000 rpm, 10 min); after BSA blocking, wash twice with PBST.

ActivationEDC/NHS activation in PBS for 30 min; BSA blocking after antibody conjugation.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	MdP particles	10.0 mg	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Other	PBS buffer	2.0 mL	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Additive	EDC	100 uL · 10.0 mg mL-1	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Additive	NHS	50.0 uL · 10.0 mg mL-1	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Other	PCT detection antibody (Ab2)	40.0 uL · 5.00 mg mL-1	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP
Other	BSA solution	2.0 mL blocking; 1.0 mL storage · 3.0%; 0.10%	SI text · 1.2 Preparation procedures of Ab1-MBs and Ab2-MdP

Complete recipeSource: Main

Route 3: Solvothermal

2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP

Metal precursorsZrCl4, 400 mg

Linker precursorsTCPP, 50.0 mg

SolventsDMF, 60 mL initially; 60 mL DMF for HCl activation dispersion

Additivesdodecanoic acid (DA), 32.0 g; hydrochloric acid, 1.30 mL of 12 M

Atmospherenot stated

Temperature120

Time36 h solvothermal growth; 12 h HCl activation

Work-upCool; centrifuge at 8000 rpm for 10 min; wash three times with DMF; after HCl activation centrifuge, wash three times with DMF.

ActivationHCl activation in DMF at 120 C for 12 h; dried at 60 C in vacuum oven for 24 h.

Scalability contextBatch solvothermal synthesis in 200 mL PTFE-lined vessel.

Show 5 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	ZrCl4	400 mg	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Linker	TCPP	50.0 mg	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Solvent	DMF	60 mL + 60 mL	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Additive	dodecanoic acid (DA)	32.0 g	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Acid	hydrochloric acid	1.30 mL · 12 M	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP

Partial recipeSource: Both

Route 4: Hydrothermal

14 · Figures and Tables · Table S2

Metal precursorsdPCN-224 nanoparticle, 40.0 mg inferred from base MdP route; MnCl2 additive, 4 mg

Linker precursorsdPCN-224 framework contains TCPP

Solventswater; ethanol wash

AdditivesNH4Cl and NH3.H2O follow base MdP route unless varied; Table S2 reports MnCl2 amount only

Atmospherenot stated

Temperature140

Time12 h hydrothermal; 10 min ultrasonication before heating

Oxidant / reductantHydrothermal conversion of MnCl2 in basic ammonium solution to Mn oxide-loaded dPCN; explicit oxidant not stated.

Work-upBase MdP route: cool; centrifuge at 8000 rpm for 10 min; wash three times with water and EtOH.

ActivationBase MdP route: dried at 60 C in a vacuum oven for 24 h.

Scalability contextSmall batch hydrothermal variant series.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	dPCN-224 nanoparticle	40.0 mg in base route	14 · Figures and Tables · Table S2
Metal Source	MnCl2	4 mg	14 · Figures and Tables · Table S2
Additive	NH4Cl	963 mg in base route	14 · Figures and Tables · Table S2
Base	NH3.H2O	700 uL in base route	14 · Figures and Tables · Table S2
Solvent	water	30.0 mL total in base route	14 · Figures and Tables · Table S2
Solvent	EtOH	wash solvent	14 · Figures and Tables · Table S2

Partial recipeSource: Both

Route 5: Hydrothermal

14 · Figures and Tables · Table S2

Metal precursorsdPCN-224 nanoparticle, 40.0 mg inferred from base MdP route; MnCl2 additive, 8 mg

Linker precursorsdPCN-224 framework contains TCPP

Solventswater; ethanol wash

AdditivesNH4Cl and NH3.H2O follow base MdP route unless varied; Table S2 reports MnCl2 amount only

Atmospherenot stated

Temperature140

Time12 h hydrothermal; 10 min ultrasonication before heating

Oxidant / reductantHydrothermal conversion of MnCl2 in basic ammonium solution to Mn oxide-loaded dPCN; explicit oxidant not stated.

Work-upBase MdP route: cool; centrifuge at 8000 rpm for 10 min; wash three times with water and EtOH.

ActivationBase MdP route: dried at 60 C in a vacuum oven for 24 h.

Scalability contextSmall batch hydrothermal variant series.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	dPCN-224 nanoparticle	40.0 mg in base route	14 · Figures and Tables · Table S2
Metal Source	MnCl2	8 mg	14 · Figures and Tables · Table S2
Additive	NH4Cl	963 mg in base route	14 · Figures and Tables · Table S2
Base	NH3.H2O	700 uL in base route	14 · Figures and Tables · Table S2
Solvent	water	30.0 mL total in base route	14 · Figures and Tables · Table S2
Solvent	EtOH	wash solvent	14 · Figures and Tables · Table S2

Partial recipeSource: Both

Route 6: Hydrothermal

14 · Figures and Tables · Table S2

Metal precursorsdPCN-224 nanoparticle, 40.0 mg inferred from base MdP route; MnCl2 additive, 12 mg

Linker precursorsdPCN-224 framework contains TCPP

Solventswater; ethanol wash

AdditivesNH4Cl and NH3.H2O follow base MdP route unless varied; Table S2 reports MnCl2 amount only

Atmospherenot stated

Temperature140

Time12 h hydrothermal; 10 min ultrasonication before heating

Oxidant / reductantHydrothermal conversion of MnCl2 in basic ammonium solution to Mn oxide-loaded dPCN; explicit oxidant not stated.

Work-upBase MdP route: cool; centrifuge at 8000 rpm for 10 min; wash three times with water and EtOH.

ActivationBase MdP route: dried at 60 C in a vacuum oven for 24 h.

Scalability contextSmall batch hydrothermal variant series.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	dPCN-224 nanoparticle	40.0 mg in base route	14 · Figures and Tables · Table S2
Metal Source	MnCl2	12 mg	14 · Figures and Tables · Table S2
Additive	NH4Cl	963 mg in base route	14 · Figures and Tables · Table S2
Base	NH3.H2O	700 uL in base route	14 · Figures and Tables · Table S2
Solvent	water	30.0 mL total in base route	14 · Figures and Tables · Table S2
Solvent	EtOH	wash solvent	14 · Figures and Tables · Table S2

Partial recipeSource: Both

Route 7: Hydrothermal

14 · Figures and Tables · Table S2

Metal precursorsdPCN-224 nanoparticle, 40.0 mg inferred from base MdP route; MnCl2 additive, 16 mg

Linker precursorsdPCN-224 framework contains TCPP

Solventswater; ethanol wash

AdditivesNH4Cl and NH3.H2O follow base MdP route unless varied; Table S2 reports MnCl2 amount only

Atmospherenot stated

Temperature140

Time12 h hydrothermal; 10 min ultrasonication before heating

Oxidant / reductantHydrothermal conversion of MnCl2 in basic ammonium solution to Mn oxide-loaded dPCN; explicit oxidant not stated.

Work-upBase MdP route: cool; centrifuge at 8000 rpm for 10 min; wash three times with water and EtOH.

ActivationBase MdP route: dried at 60 C in a vacuum oven for 24 h.

Scalability contextSmall batch hydrothermal variant series.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	dPCN-224 nanoparticle	40.0 mg in base route	14 · Figures and Tables · Table S2
Metal Source	MnCl2	16 mg	14 · Figures and Tables · Table S2
Additive	NH4Cl	963 mg in base route	14 · Figures and Tables · Table S2
Base	NH3.H2O	700 uL in base route	14 · Figures and Tables · Table S2
Solvent	water	30.0 mL total in base route	14 · Figures and Tables · Table S2
Solvent	EtOH	wash solvent	14 · Figures and Tables · Table S2

Partial recipeSource: Both

Route 8: Hydrothermal

14 · Figures and Tables · Table S2

Metal precursorsdPCN-224 nanoparticle, 40.0 mg inferred from base MdP route; MnCl2 additive, 20 mg

Linker precursorsdPCN-224 framework contains TCPP

Solventswater; ethanol wash

AdditivesNH4Cl and NH3.H2O follow base MdP route unless varied; Table S2 reports MnCl2 amount only

Atmospherenot stated

Temperature140

Time12 h hydrothermal; 10 min ultrasonication before heating

Oxidant / reductantHydrothermal conversion of MnCl2 in basic ammonium solution to Mn oxide-loaded dPCN; explicit oxidant not stated.

Work-upBase MdP route: cool; centrifuge at 8000 rpm for 10 min; wash three times with water and EtOH.

ActivationBase MdP route: dried at 60 C in a vacuum oven for 24 h.

Scalability contextSmall batch hydrothermal variant series.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	dPCN-224 nanoparticle	40.0 mg in base route	14 · Figures and Tables · Table S2
Metal Source	MnCl2	20 mg	14 · Figures and Tables · Table S2
Additive	NH4Cl	963 mg in base route	14 · Figures and Tables · Table S2
Base	NH3.H2O	700 uL in base route	14 · Figures and Tables · Table S2
Solvent	water	30.0 mL total in base route	14 · Figures and Tables · Table S2
Solvent	EtOH	wash solvent	14 · Figures and Tables · Table S2

Partial recipeSource: Both

Route 9: Hydrothermal

14 · Figures and Tables · Table S2

Metal precursorsdPCN-224 nanoparticle, 40.0 mg inferred from base MdP route; MnCl2 additive, 24 mg

Linker precursorsdPCN-224 framework contains TCPP

Solventswater; ethanol wash

AdditivesNH4Cl and NH3.H2O follow base MdP route unless varied; Table S2 reports MnCl2 amount only

Atmospherenot stated

Temperature140

Time12 h hydrothermal; 10 min ultrasonication before heating

Oxidant / reductantHydrothermal conversion of MnCl2 in basic ammonium solution to Mn oxide-loaded dPCN; explicit oxidant not stated.

Work-upBase MdP route: cool; centrifuge at 8000 rpm for 10 min; wash three times with water and EtOH.

ActivationBase MdP route: dried at 60 C in a vacuum oven for 24 h.

Scalability contextSmall batch hydrothermal variant series.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	dPCN-224 nanoparticle	40.0 mg in base route	14 · Figures and Tables · Table S2
Metal Source	MnCl2	24 mg	14 · Figures and Tables · Table S2
Additive	NH4Cl	963 mg in base route	14 · Figures and Tables · Table S2
Base	NH3.H2O	700 uL in base route	14 · Figures and Tables · Table S2
Solvent	water	30.0 mL total in base route	14 · Figures and Tables · Table S2
Solvent	EtOH	wash solvent	14 · Figures and Tables · Table S2

Complete recipeSource: Main

Route 10: Hydrothermal

2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP

Metal precursorsMnCl2, 20.0 mg; dPCN nanoparticle, 40.0 mg

Linker precursorsdPCN-224 framework contains TCPP

Solventswater, 15.0 mL for dPCN dispersion and 15.0 mL for Mn salt solution; ethanol for washing

AdditivesNH4Cl, 963 mg; NH3.H2O, 700 uL

Atmospherenot stated

Temperature140

Time12 h hydrothermal; 10 min ultrasonication before heating

Oxidant / reductantDissolved MnCl2 converted in basic ammonium solution to mixed-valence Mn oxides during hydrothermal treatment; explicit oxidant not stated.

Work-upCool; centrifuge at 8000 rpm for 10 min; wash three times with water and EtOH.

ActivationDried at 60 C in a vacuum oven for 24 h.

Scalability contextBatch hydrothermal loading in 100 mL PTFE-lined vessel.

Show 6 structured reagent records

Role	Reagent	Amount / concentration	Source
Other	dPCN nanoparticle	40.0 mg	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Metal Source	MnCl2	20.0 mg	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Additive	NH4Cl	963 mg	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Base	NH3.H2O	700 uL	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Solvent	water	30.0 mL total	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Solvent	EtOH	wash solvent	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP

Partial recipeSource: Main

Route 11: Solvothermal

2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP

Metal precursorsZrCl4, 400 mg

Linker precursorsTCPP, 50.0 mg

SolventsDMF, 60 mL

Additivesbenzoic acid (BA), 8.00 g

Atmospherenot stated

Temperature120

Time36 h growth; follow-on activation inferred from dPCN procedure

Work-upAuthors state following procedures were the same as dPCN-224 preparation.

ActivationSame as dPCN-224 procedure according to main text; exact repetition not restated.

Scalability contextBatch solvothermal synthesis.

Show 4 structured reagent records

Role	Reagent	Amount / concentration	Source
Metal Source	ZrCl4	400 mg	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Linker	TCPP	50.0 mg	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Solvent	DMF	60 mL	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP
Additive	benzoic acid (BA)	8.00 g	2 · 2.2 Preparation of PCN-224, dPCN-224 and MdP