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

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns 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 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	20	2,906.8	205.2	7.1
2	20	702.3	57.6	8.2
3	20	155.7	13.9	8.9

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	3,039.6	237.3	7.8
2	24	712.1	40.6	5.7
3	24	182.3	10.8	5.9

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 serum	1:16	105	94-111
Cell culture supernatant	1:8	87	81-94
Saliva	1:16	102	93-107

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 serum	Cell culture supernatant	Saliva
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	106	115	101
	Range (%)	97-117	111-120	95-108
1:8	Average% of Expected	86	116	112
	Range (%)	79-90	109-124	106-122
1:16	Average% of Expected	79	126	121
	Range (%)	70-93	122-130	120-122

CALIBRATION

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