DESCRIPTION

Fibrillin 1 (also known as FBN1) belongs to the fibrillin family, which includes FBN1, FBN2, and FBN3. Fibrillin 1 is a large extracellular matrix protein that assembles into microfibrils that fulfill various structural and instructive roles. Fibrillin 1 comprises the core of fibrillin-containing microfibrils, which provide the principal scaffold for elastic fiber and lamellae formation required for elastic tissue properties. Fibrillin1 is ubiquitously present in many tissues throughout the body. Mutations in the Fibrillin 1 gene can lead to a variety of genetic disorders, such as Marfan syndrome.

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	2,066.5	70.6	3.4
2	8	514.5	21.0	4.1
3	8	255.0	11.0	4.3

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,062.2	69.8	3.4
2	16	515.5	28.7	5.6
3	16	258.6	10.8	4.2

RECOVERY

The recovery of human Fibrillin 1 spiked to three different levels throughout the range of the assay in human plasma was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:40	81	71-87

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. (The human serum was initially diluted 1:5. )

		Human plasma
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	98
	Range (%)	96-101
1:8	Average% of Expected	89
	Range (%)	83-95
1:16	Average% of Expected	78
	Range (%)	76-80

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Fibrillin 1 produced at Sunlong Medical™.