Tissue Transglutaminase Ab IgA ELISA Kit (tTG)

Full Name: Tissue Transglutaminase Ab IgA ELISA Kit (tTG)
Reactivity: Human
Sample Type: Serum, Plasma
Sensitivity: 1.0 U/ml

BACKGROUND

Tissue transglutaminase (tTG) is a multifaceted enzyme capable of post-translationally modifying proteins through transamidation reactions that form covalent bonds between lysine and glutamine amino acid residues. This crosslinking of proteins by tTG can profoundly alter their structural properties and biological functions. tTG is expressed ubiquitously in mammalian tissues, with the highest levels found intracellularly in the cytosol.

The structure of tTG consists of four domains: an N-terminal β-sandwich domain that anchors the enzyme to fibronectin in the extracellular matrix, a catalytic core domain housing the active site, and two C-terminal β-barrel domains regulating access to the active site. The active site contains a key cysteine residue essential for catalyzing transamidation reactions. Binding of calcium ions induces conformational changes in tTG that enhance active site accessibility and activate its crosslinking function.

One well-understood role of tTG is in the pathogenesis of celiac disease. In the intestine, tTG deamidates gluten peptides by converting glutamine amino acids to glutamate. This increases gluten peptide binding affinity for HLA-DQ2/8 molecules on antigen presenting cells. Display of deamidated gluten peptides triggers activation of inflammatory CD4+ T cells specific for gluten, leading to intestinal damage and clinical manifestations of celiac disease. tTG autoantibodies are produced in celiac disease and utilized clinically to aid diagnosis.

Apart from celiac disease, abnormal expression and activity of tTG have been linked to numerous other disease processes like fibrosis, cancer, neurodegeneration, thrombosis, and autoimmunity. For instance, excessive crosslinking by tTG can promote extracellular matrix accumulation in fibrotic disorders.

INTENDED USE

Human tissue transglutaminase Ab IgA ELISA kit is a procedure intended for in-vitro quantitative measurement of IgA class auto-antibodies direct to tissue transglutaminase (tTG, TG2-IgA, anti-tissue transglutaminase, tissue transglutaminase IgA) in human serum and plasma samples. 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, recombinant tTG antibodies bound to microwells.
Calibrators (A-F): Concentration 0, 5, 10, 25, 75, 200 U/ml, made up from tissue transglutaminase antibody.
Controls Positive and Control Negative: Containing tissue transglutaminase antibody.
Sample Buffer P (5x).
Enzyme Conjugate: Contains anti-human IgA antibodies, HRP labelled.
TMB Substrate.
Stop Solution: Contains acid.
Wash Buffer (50x Concentrated).
Instruction Manual.
Certificate of Analysis.

SENSITIVITY

The minimum detection sensitivity level of tissue transglutaminase IgA autoantibodies (tissue transglutaminase Ab IgA, tTG IgA, TG2 IgA) using current tissue transglutaminase IgA ELISA kit was 1.0 U/ml. The dynamic range for this assay is 5.0 – 200.0 U/ml.

ASSAY CHARACTERISTICS

– Measuring Range: 0 – 200 U/ml
– Expected Values: Cut-off 10 U/ml.
– Results Interpretation: Negative <10 U/ml, Positive ≥ 10 U/ml.
– Limit Of Detection (LOD): Functional sensitivity was 1 U/ml.
– Linearity: 94 – 105%
– Intra Assay: 4.4 – 10.4%
– Inter Assay: 10.6 – 13.7%
– Interference: None.
– Clinical Diagnosis: Sensitivity (100.0%), Specificity (97.8%), Overall agreement (98.4%).

REFERENCES

IgA anti-tissue transglutaminase: setting the stage for coeliac disease screening. Eur J Gastroenterol Hepatol. (2001) 13 (6): 635-7. Review. Schuppan D. and Hahn E.G.
Overview of biomarkers for diagnosis and monitoring of celiac disease. Adv Clin Chem. (2015) 68: 1-55. Review. Brusca I.
Anti-tissue transglutaminase antibodies and their role in the investigation of coeliac disease. Ann Clin Biochem. (2006) 43 (Pt 2): 105-17. Review. Hill P.G. and McMillan S.A.

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