DESCRIPTION

Matrix metalloproteinase 3 (MMP-3), also known as Stromelysin-1, localized to chromosome 11q22.3, is a member of the metalloproteinase enzyme family, which consists of approximately 20 zinc-dependent endopeptidases. The main role of MMP3 is involved in the breakdown of the extracellular matrix (ECM) proteins during tissue remodeling in normal physiological processes, such as embryonic development and reproduction. The early studies on MMP-3 focused on its roles in collagen degradation and various pathological conditions, including arthritis, fibrosis, and cancer.

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	1,216.5	22.2	1.8
2	8	299.4	6.5	2.2
3	8	157.8	4.9	3.1

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,201.8	28.3	2.4
2	16	302.5	14.8	4.9
3	16	158.5	5.0	3.1

RECOVERY

The recovery of human MMP3 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:40	106	99-112
	1:80	102	95-107
Cell culture supernatant	1:64	104	93-116
	1:128	102	98-107

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. (The human plasma was initially diluted 1:10. The cell culture supernatant was initially diluted 1:8.)

		Human plasma	Cell culture supernatant
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	116	97
	Range (%)	115-117	92-101
1:8	Average% of Expected	111	110
	Range (%)	102-119	105-115
1:16	Average% of Expected	110	102
	Range (%)	104-117	102-103

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human MMP3 produced at Sunlong Medical™.