Human ANCA-C ELISA kit is designed for measuring IgG autoantibodies to proteinase 3 (PR3, anti-PR3, ANCA-C, cytoplasmic anti-neutrophil cytoplasmic antibodies, c-ANCA) using human serum or plasma. This assay has a minimum analytical sensitivity limit of 0.5 U/ml.
Cytoplasmic anti-neutrophil cytoplasmic antibodies (cANCA, ANCA-C or ANCA-PR3) are autoantibodies produced by the body to attack its own proteins. Proteinase 3 (PR3) is one of the most common target proteins that is found to occur throughout the neutrophil cytoplasm. They are mainly present as granulomatosis patients with polyangiitis (this is present in over 90%). It is able to catabolise many human proteins for example elastin, four subclasses of IgG and proteoglycans.
There are scientific publications that indicate specific IgG types of ANCA that are directed against PR3 of monocytes and neutrophils are instrumental in the vascular damage (especially pathophysiology) which leads to the excessive activation of neutrophils and in many cases the destruction of vessel walls. There are many animal models and in vitro studies that indicate a pathogenic role for it through its ability to induce many inflammatory reactions.
ANCA-PR3 is regarded as a biomarker for eosinophilic, granulomatosis and microscopic polyangiitis where it is displaying a high specificity. There are many factors (for example glycosylation and paratope) that are found to influence the pathogenicity, in order to develop new therapies and even biomarkers then all these factors will need to be addressed. The following ELISA method can be used to interpret samples that are difficult to analyse by indirect immunofluorescence techniques such as those with high background fluorescence or those which exhibit several antibodies simultaneously.
All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.
- Divisible Microplate: Highly purified PR3 is bound to microwells.
- Calibrator A-F: Concentration 0, 5, 10, 20, 40, 100 U/ml.
- Control Positive And Control Negative: Antibodies in a serum/buffer matrix.
- Sample Buffer (5x).
- Enzyme Conjugate: Containing anti-human IgG antibodies, HRP labelled.
- Substrate: Tetramethylbenzidin (TMB).
- Stop Solution.
- Wash Buffer (50x).
- Instruction For Use.
- Certificate Of Analysis.
The minimum detection sensitivity level of human IgG autoantibodies to proteinase 3 (anti-PR3, PR3, ANCA-C, c-ANCA, cytoplasmic anti-neutrophil cytoplasmic antibodies) using current ANCA-C ELISA kit was 0.5 U/ml. The dynamic range for this assay is 5 – 100.0 U/ml.
– Measuring Range: 0 – 100 U/ml
– Expected Values: In a normal range = Cut-off Index 5.0 U/ml, Positive: Index ≥ 5.0 U/ml, Negative: Index < 5.0 U/ml.
– Linearity: 96 – 108%
– Intra Assay Precision: 2.2 – 4.7%
– Inter Assay Precision: 3.9 – 8.8%
– Clinical Diagnosis: Sensitivity (85.2%), Specificity (98.8%), Overall agreement (96.1%).
- Human proteinase 3 (PR3) and its binding molecules: implications for inflammatory and PR3-related autoimmune responses. Ann N Y Acad Sci. (2007) 1109: 84-92. Müller A., et al.
- PR3-ANCA: a promising biomarker for ulcerative colitis with extensive disease. Clin Chim Acta. (2013) 424: 267-73. Mahler M., et al.
- Antibodies directed against neutrophils (C-ANCA and P-ANCA) are of distinct diagnostic value in systemic vasculitis. Q J Med. (1990) 77 (284): 1287-96. Venning M.C., et al.
- Performance evaluation of three assays for the detection of PR3-ANCA in granulomatosis with polyangiitis in daily practice. Autoimmun Rev. (2013) 12 (12): 1118-22. Noel N, André C., et al.
- Full Name: ANCA-C ELISA Kit (Proteinase 3, PR3)
- Reactivity: Human
- Sample Type: Serum, Plasma
- Sensitivity: 0.5 U/ml
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