GBM Ab ELISA Kit (Glomerular Basement Membrane)

Full Name: GBM Ab ELISA Kit (Glomerular Basement Membrane)
Reactivity: Human
Sample Type: Plasma, Serum
Sensitivity: 1.0 U/ml

BACKGROUND

Glomerular basement membrane (GBM) is the basal lamina layer of glomerulus which can be found in the kidney. The GBM, glomerular capillary endothelial cells and filtration slits (between podocytes) all are vital in performing the function of filtrating the glomerulus. This is a vital process which is responsible for separating the blood present in the capillaries from the filtrate (formed in the Bowman’s capsule). It contains three layers: the lamina rara externa which is adjacent to podocyte processes, lamina densa which is the dark central zone and the lamina rara interna which is adjacent to endothelial cells. It is also made up of mesangial cells and modified pericytes which can separate capillaries from each other in other parts of the body. The podocytes which are responsible for adjoining them contain filtration slits with a diameter 25nm (formed by pseudopodia).

The pathological condition which that associated include: Goodpasture’s syndrome (which can also be called anti-GBM disease), Nephrotic syndrome, diabetic glomerulosclerosis and Alport syndrome. Goodpasture’s syndrome is a condition that is affecting the lungs and the kidneys, if it is left untreated it can lead to serious complications for example: respiratory failure due to severe bleeding within the lungs or kidneys failure due to severe kidney inflammation.

INTENDED USE

Human GBM Ab ELISA kit can be used for in-vitro quantitative analysis of anti IgG auto-antibodies against glomerular basement membrane (GBM IgG, GBM Ab) in human plasma and serum. This assay has a minimum analytical sensitivity limit of 1.0 U/ml.

CONTENT

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: Consisting of 12 modules of 8 wells, highly purified anti glomerular basement membrane is bound to microwells.
• Calibrator A-E: Concentrations 0, 20, 40, 80, 200 U/ml, made up from human GBM antibody.
• Control Positive and Control Negative: Containing human GBM Ab in a serum/buffer matrix.
• Sample Buffer P (5x).
• Enzyme Conjugate: Contains anti-human IgG antibodies, HRP labelled.
• TMB Substrate.
• Stop Solution: Contains acid.
• Wash Buffer (50x).
• Instruction for Use.
• Certificate of Analysis.

SENSITIVITY

The minimum detection sensitivity level of IgG antibody to human glomerular basement membrane (GBM-IgG, GBM Ab) using current GBM Ab ELISA kit was 1.0 U/ml. The dynamic range for this assay is 20.0 – 200.0 U/ml.

ASSAY CHARACTERISTICS

– Measuring Range: 0 – 200 U/ml.
– Expected Values: Samples from healthy blood donors displayed a cut-off Index 20 U/ml.
– Results Interpretation: Negative: < 20 U/ml, Positive: ≥ 20 U/ml
– Linearity: 93 – 101%
– Limit Of Detection (LOD): Functional sensitivity was determined to be 1 U/ml.
– Intra Assay Precision: 5.3 – 5.7%
– Inter Assay Precision: 5.4 – 6.4%
– Interfering Substances: None.
– Clinical Diagnosis: Sensitivity (100.0%), Specificity (99.3%), Overall agreement (99.4%).

REFERENCES

1. Ultrastructure of developing kidney glomerular basement membranes: temporal changes in binding of anti-laminin IgG and cationized ferritin. Microsc Res Tech. (1994) 28 (2): 81-94. Review. Abrahamson D.R. and St John P.L.
2. Organogenesis of the kidney glomerulus: focus on the glomerular basement membrane. Organogenesis. (2011) 7 (2): 75-82. Review. Miner J.H.
3. Goodpasture’s syndrome: a clinical update. Autoimmun Rev. (2015) 14 (3): 246-53. Review. Greco A., et al.
4. Concurrent anti-glomerular basement membrane disease and membranous glomerulonephritis: a case report and literature review. Clin Nephrol. (2006) 66 (2): 120-7. Review. Troxell M.L., et al.
5. Diabetic nephropathy with interstitial nephritis presenting with a false-positive anti-GBM antibody. Clin Nephrol. (2002) 57 (5): 381-5. DeAangelo A.J., et al.

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