DESCRIPTION

Matrix metalloproteinase 1 (MMP1), which is also known as interstitial collagenase and fibroblast collagenase is a member of the matrix metalloproteinases (MMPs) family. Proteins in this family mainly participate in the breakdown of extracellular matrix both in normal physiological processes and disease processes. Several factors contribute to the expression of MMP1, including endogenous factors, such as polymorphisms and epigenetic regulation in the promoter region of MMP1, as well as exogenous factors, such as the tumor microenvironment. A variety of MMP1 promoter genotypic polymorphisms have been reported in various ethnic populations, and their contributions to different disease risks have been explored. In the tumor microenvironment, a variety of inflammatory factors, including interleukin-8 (IL-8), IL-1β and tumor necrosis factor-α (TNF-α), have been reported to induce MMP1 expression in cancer cells.

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	981.3	14.2	1.4
2	8	258.6	6.1	2.4
3	8	129.3	2.4	1.9

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	972.5	19.6	2.0
2	16	255.5	6.7	2.6
3	16	128.2	3.0	2.3

RECOVERY

The recovery of human MMP-1 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:32	85	76-91
Human plasma	1:16	95	81-113
Cell culture supernatant	1:16	98	82-118

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 1:4.)

		Human serum	Human plasma	Cell culturesupernatant
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	101	92	88
	Range (%)	85-117	90-94	78-99
1:8	Average% of Expected	111	88	91
	Range (%)	110-113	81-95	71-111
1:16	Average% of Expected	96	80	94
	Range (%)	84-108	78-81	74-114

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human MMP1 produced at Sunlong Medical™.