title: Development of a sandwich ELISA assay for quantification of human tissue transglutaminase in cell lysates and tissue homogenates.
abstract: 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.
authors.affiliation: Research Department, Covalab S.A.S, 11 Avenue Albert Einstein, 69100, Villeurbanne, France.
authors.forename: Éva
authors.initials: É
authors.lastname: Sivadó
authors.affiliation: Research Department, Covalab S.A.S, 11 Avenue Albert Einstein, 69100, Villeurbanne, France.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Lareure
authors.affiliation: Research Department, Covalab S.A.S, 11 Avenue Albert Einstein, 69100, Villeurbanne, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Attuil-Audenis
authors.affiliation: Research Department, Covalab S.A.S, 11 Avenue Albert Einstein, 69100, Villeurbanne, France.
authors.forename: Saïd El
authors.initials: SE
authors.lastname: Alaoui
authors.affiliation: Research Department, Covalab S.A.S, 11 Avenue Albert Einstein, 69100, Villeurbanne, France. vthomas@covalab.com.
authors.forename: Vincent
authors.initials: V
authors.lastname: Thomas
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards*; D005260:Female; D019204:GTP-Binding Proteins / Q000302:isolation & purification*; D006168:Guinea Pigs; D057809:HEK293 Cells; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification; D008099:Liver / Q000737:chemistry / Q000201:enzymology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000737:chemistry / Q000201:enzymology*; D015588:Observer Variation; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D011503:Transglutaminases / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins
keywords: Monoclonal antibody; Polyclonal antibody; Sandwich ELISA; Tissue transglutaminase