DESCRIPTION

MPO(myeloperoxidase) is a peroxidase enzyme presented in the azurophilic granules of polymorphonuclear (PMN) leukocytes and monocytes. Plasma concentration of MPO can be used as a specific marker of PMN activation. MPO catalyzes the production of hypochlorous acid (HClO) from hydrogen peroxide (H2O2) and chloride anion (Cl-, or the equivalent from a non-chlorine halide). This enzymatic system plays an important role in human defense against microorganisms. The serum/plasma MPO levels have been associated with a variety of clinical conditions including inflammatory diseases, atherosclerosis, ischaemic stroke, hypertension, heart failure, risk of cardiovascular events and so on.

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	1,309.5	67.3	5.1
2	20	307.7	15.5	5.0
3	20	70.7	4.5	6.4

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	1,267.8	35.5	2.8
2	24	299.5	20.8	6.9
3	24	70.6	5.9	8.3

RECOVERY

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

Sample Type		Average% of Expected	Range (%)
Human serum	1:800	85	77-92
	1:1,600	87	78-93
Cell culture supernatant	1:160	78	71-83
	1:320	83	77-98
Urine	1:16	79	74-92
	1:32	79	72-87
Saliva	1:800	113	105-120
	1:1,600	87	79-94
Cell lysate	1:320,000	92	72-114
	1:640,000	100	87-110

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 1:100. The cell culture supernatant was initially diluted 1:20. The urine was initially diluted 1:2. The saliva was initially diluted 1:100. The cell lysate was initially diluted 1:40,000.)

		Human serum	Cell culturesupernatant	Urine	Saliva	Cell lysate
1:2	Average% ofExpected	100	100	100	100	100
	Range (%)	-	-	-	-	-
1:4	Average% ofExpected	109	107	110	96	103
	Range (%)	108-111	103-112	103-117	95-96	102-103
1:8	Average% ofExpected	113	105	117	100	109
	Range (%)	112-113	87-119	105-118	97-102	102-116
1:16	Average% ofExpected	118	109	121	99	112
	Range (%)	116-120	90-124	111-131	98-100	104-121

CALIBRATION

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

Ordering & shipping

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