Comparison of ELISA- and SIMOA-based quantification of plasma Aβ ratios for early detection of cerebral amyloidosis

De Meyer, Steffi; Schaeverbeke, Jolien M.; Verberk, Inge M. W.; Gille, Benjamin; De Schaepdryver, Maxim; Luckett, Emma S.; Gabel, Silvy; Bruffaerts, Rose; Mauroo, Kimberley; Thijssen, Elisabeth H.; Stoops, Erik; Vanderstichele, Hugo M.; Teunissen, Charlotte E.; Vandenberghe, Rik; Poesen, Koen

doi:10.1186/s13195-020-00728-w

Alzheimer's Research & Therapy

Table 1 Analytical assay characteristics

From: Comparison of ELISA- and SIMOA-based quantification of plasma Aβ ratios for early detection of cerebral amyloidosis

	Platform	ELISA colorimetric			SIMOA
	Analyte	Aβ_1–42	Aβ_1–40	t-tau	Aβ_1–42	Aβ_1–40
Assay	Provider	EUROIMMUN	EUROIMMUN	ADx	ADx	ADx
	Catalogue number	EQ 6521-9601	EQ 6511-9601	NA	NA	NA
	Biofluid	EDTA plasma	EDTA plasma	EDTA plasma	EDTA plasma	EDTA plasma
	Status	Commercial	Commercial	Prototype	Prototype	Prototype
	Specificity	Aβ_1–42	Aβ_1–40	6 tau isoforms	Aβ_1–42	Aβ_1–40
Dilution	Pre-dilution factor^a	4	4	4	4	20
Dilution	Final sample dilution^b	5	5	5	5.8	29
Calibrator	Type	Recombinant	Recombinant	Recombinant	Recombinant	Recombinant
	No. of calibrator points	7	7	7	7	7
	Range, pg/mL	1–40	1–75	1–100	1–64	1–64
Patient samples	Number	199	199	199	199	199
	Range, pg/mL	8.1–57.1	22.4–311.8	15.0–102.1	7.9–44.0	60.7–160.0
	Within cal. range, %	100%	100%	100%	100%	100%
CV%	Intra-assay conc	1.63	1.67	2.15	4.33	2.24
CV%	Inter-assay conc	4.82	2.58	13.55	8.59	5.95
Analytical sensitivity	LoD, pg/mL	2.4	5.7	Not determined	0.46	0.77
	LoQ, pg/mL	3.5	12.5	Not determined	1.21	1.57
	S/N ratio	7.60	65.4	Not determined	11.0	240
Antibodies [26, 27]	Name capture	ADx102 (21F12)	ADx103 (2G3)	ADx203	ADx102 (21F12)	ADx103 (2G3)
	Epitope capture (AA)	Aβ_34–42	Aβ_33–40	Tau_194–204	Aβ_34–42	Aβ_33–40
	Name detector	2G3	2G3	ADx204	2G3	2G3
	Epitope detector (AA)	Aβ_1–5	Aβ_1–5	Tau_N-terminus	Aβ_1–5	Aβ_1–5
Assay protocol	Incubation times, h	3–0.5–0.5	3–0.5–0.5	3–0.5–0.5	2–0.08	2–0.08
	Incubation T, °C	18–25	18–25	18–25	18–25	18–25
	Curve fit	4PL	4PL	4PL	4PL	4PL
QC panel, pg/mL	QC1: high	30.5	189.2	22.3	13.7	83.2
	QC2: intermediate	26.6	149.1	18.9	19.7	75.9
	QC3: intermediate	22.0	116.8	21.7	13.4	69.7
	QC4: low	20.7	107.3	23.7	10.5	33.3
	QC5: low spiked	14.4	0.0	NA	21.6	21.8
	QC6: high spiked	117.9	125.6	NA	173.7	181.2
	C1: low kit control	23.3	114.6	NA	NA	NA
	C2: high kit control	45.1	186.9	NA	NA	NA

AA amino acid, Aβ β-amyloid, CV coefficient of variation, LoD limit of detection, LoQ limit of quantification, S/N ratio signal to noise ratio, QC quality control, T temperature, t-tau total tau
^aSample pre-dilution was performed using assay diluent in polypropylene low-binding 96-well microplates
^bFinal sample dilution during sample incubation step in assay protocol

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ISSN: 1758-9193

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