DESCRIPTION

Vascular endothelial growth factor C (VEGF-C) is a protein that is a member of the platelet-derived growth factor / vascular endothelial growth factor (PDGF/VEGF) family. The main function of VEGF-C is to promote the growth of lymphatic vessels (lymphangiogenesis). It acts on lymphatic endothelial cells (LECs) primarily via its receptor VEGFR-3 promoting survival, growth and migration. Apart from vascular targets, VEGF-C is also important for neural development and blood pressure regulation.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	5,859.8	212.0	3.6
2	8	264.3	15.6	5.9
3	8	23.5	1.7	7.3

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	6,264.0	622.5	9.9
2	16	278.4	22.0	7.9
3	16	23.7	1.6	6.7

RECOVERY

The recovery of human VEGF-C spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:4	98	93-104
	1:8	105	103-108
Cell culture supernatant	1:8	98	94-100
	1:16	101	101-102
Saliva	1:8	105	103-107
	1:16	96	94-98

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.

		Human plasma	Cell culturesupernatant	Saliva
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	104	92	104
	Range (%)	99-109	92-93	94-113
1:8	Average% of Expected	104	91	99
	Range (%)	87-121	90-91	91-108
1:16	Average% of Expected	101	90	88
	Range (%)	74-128	89-91	84-91

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human VEGF-C produced at Sunlong Medical™.