DESCRIPTION

CEACAM1, also known as BGP1 or CD66a, belongs to the CEA-related cell-adhesion molecule (CEACAM) subfamily, which is a part of the immunoglobulin superfamily. It plays a role in cell-cell adhesion, regulation of cell proliferation, insulin resistance, obesity, and vascular homeostasis. CEACAM1 is expressed by certain epithelial, endothelial, lymphoid, and myeloid cells. It is found on the surface of various tumors and is associated with tumor progression, metastasis and poor prognosis. CEACAM1 serves as a specific biomarker for Non-small cell lung cancer (NSCLC) and can be used for diagnosis and prognosis of melanoma.

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	1,209.8	25.7	2.1
2	8	289.6	4.4	1.5
3	8	155.5	4.2	2.7

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,220.1	25.9	2.1
2	16	291.9	6.8	2.3
3	16	154.8	3.9	2.5

RECOVERY

The recovery of human CEACAM1/CD66a 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	89	85-92
	1:32	98	90-104
Cell culture supernatant	1:2	100	96-104
	1:4	106	103-108

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
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	115	81
	Range (%)	111-118	72-88
1:8	Average% of Expected	101	99
	Range (%)	96-106	89-111
1:16	Average% of Expected	84	103
	Range (%)	77-91	91-118

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human CEACAM1/CD66a One-Step ELISA Kit produced at Sunlong Medical™.