Insulin Ab ELISA Kit

Full Name: Insulin Ab ELISA Kit
Reactivity: Human
Sample Type: Serum, Plasma
Sensitivity: 0.5 U/ml

BACKGROUND

Diabetes-related auto-antibodies are found to be linked to type 1 diabetes and this is a condition which is characterised by limited or in some cases fully missing secretion of insulin. Many morphological investigations have demonstrated that this process reflects the destruction of beta cells, leading to the loss of beta cell function and thereby causing inadequate production of insulin.

Auto-antibodies are present between 50-70% of children having diabetes mellitus and between 20-30% in adults. There are also instances where patients of insulin-dependent diabetes can generate antibodies targeted against the exogenous insulin. This will result in insulin-resistance and during insulin-therapy requires constant monitoring.

These auto-antibodies are causing insulin depot formation which is able to simulate a resistance against any animal insulin hormonal treatment. IgG antibodies are the most common while IgE antibodies have been found to result in causing insulin-antibody allergy, at high titers. The development of antibodies is largely dependent of the molecular structure, purity and storage conditions of the insulin administered as well as patient factors (such as age, HLA type, delivery route).

INTENDED USE

Human insulin Ab ELISA kit is a protocol for analysing in-vitro quantitative levels of IgG antibodies to insulin (anti-insulin IgG, insulin Ab, anti-insulin) in human serum and plasma. This assay has a minimum analytical sensitivity limit of 0.5 U/ml.

Mixture of highly purified preparations of porcine, bovine and recombinant human insulin are bound to the microwells.

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 mixture of bovine, porcine and recombinant human insulin is bound to microwells.
• Calibrator A-F: Concentrations 0, 6.3, 12.5, 25, 50, 100 U/ml, made up from anti-insulin antibodies in a serum/buffer matrix.
• Control Positive and Control Negative: Containing human anti-insulin antibody.
• Sample Buffer P (5x).
• Enzyme Conjugate: Contains anti-human IgG antibodies, HRP labelled.
• TMB Substrate.
• Stop Solution.
• Wash Buffer (50x).
• Instruction For Use.
• Certificate Of Analysis.

SENSITIVITY

The minimum detection sensitivity level of IgG antibodies directed against human anti-insulin (insulin Ab, anti-insulin IgG) using current insulin Ab ELISA kit was 0.5 U/ml. The dynamic range for this assay is 6.3 – 100.0 U/ml.

ASSAY CHARACTERISTICS

– Measuring Range: 0 – 100 U/ml
– Expected Values: Cut-Off 10 U/ml
– Results Interpretation: Negative: < 10 U/ml, Positive: ≥ 10 U/ml
– Linearity: 84 – 109%
– Limit Of Detection (LOD): 0.5 U/ml
– Intra Assay Precision: 2.5 – 4.0%
– Inter Assay Precision: 4.3 – 6.0%
– Interfering Substances: None
– Clinical Diagnosis: Sensitivity (72.0%), Specificity (98.8%), Overall agreement (88.5%).

REFERENCES

1. The anti-insulin trimolecular complex in type 1 diabetes. Curr Opin Endocrinol Diabetes Obes. (2010) 17 (4): 329-34. Review. Michels A.W. and Nakayama M.
2. Autoimmune insulin syndrome. Biomed Pharmacother. (1989) 43 (8): 581-6. Review. Archambeaud-Mouveroux F., et al.
3. Insulin assays: previously known and new analytical features. Clin Lab. (2003) 49 (3-4): 113-21. Review. Sapin R.
4. Role of insulin in glucose-stimulated insulin secretion in beta cells. Curr Diabetes Rev. (2005) 1 (3): 309-30. Review. Goren H.J.
5. Allergic reactions to human-insulin: a review of current knowledge and treatment options. Endocrine. (2010) 37 (1): 33-9. Review. Akinci B., et al.

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