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Development of a sandwich ELISA assay for quantification of human tissue transglutaminase in cell lysates and tissue homogenates

Sivadó, Éva, Lareure, Sabrina, Attuil-Audenis, Valérie, Alaoui, Saïd El, Thomas, Vincent
Amino acids 2017 v.49 no.3 pp. 597-604
celiac disease, cross reaction, crosslinking, detection limit, enzyme-linked immunosorbent assay, humans, monoclonal antibodies, neoplasms, neurodegenerative diseases, pathological processes and conditions, polyclonal antibodies, protein-glutamine gamma-glutamyltransferase
Tissue transglutaminase (tTG) belongs to the multigene transglutaminase family of Ca²⁺-dependent protein cross-linking enzymes. There is a strong evidence that tTG is involved in pathology, such as neurodegenerative diseases, cancer, and celiac disease. To study physiopathological implication of tTG, a sandwich immunoassay has been developed with a new monoclonal antibody for the capture and polyclonal antibody both generated in house. Using this ready to use assay, the tTG protein level can be measured in human tissue homogenates and cells extracts easily in about 4 h. The limit of detection is 1.7 ng/ml; the coefficients of intra- and inter-assay variations range from 1 to 2 % and from 7 to 10 %, respectively. The assay is specific to tTG, and no cross reactivity with TG1, TG3, TG6, TG7, or factor XIIIa was observed. Finally, in the addition to the tTG activity assay previously developed, this assay should be a valuable tool to increase our knowledge of the tTG involvement in physiological and pathological states.