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 Ca-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.

Linker-Preserved Iron Metal–Organic Framework-Based Lateral Flow Assay for Sensitive Transglutaminase 2 Detection in Urine Through Machine Learning-Assisted Colorimetric Analysis

“…(f) Comparison of the LOD and the assay time of our LFA method with those of other literature work. References: P1, P2, E1, E2, EC1, EC2, EC3, EC4, and F1 Table S10 summarizes the detailed comparison for this.…”

“…Therefore, using the equation obtained from this linear regression, the measured intensity (y) of the desalted UN and UKD patient samples was fitted to estimate the TGM2 concentration (x). Accordingly, the y values from the desalted UN-1, UN-2, UN-3, UKD-1, UKD-2, UKD-3, and 35 E2, 36 EC1, 37 EC2, 38 EC3, 39 EC4, 31 and F1. 40 UKD-4 samples were inserted into the linear equation of y = 20.93x + 0.734 (R 2 = 0.994), which resulted in the following native TGM2 concentration (x) for each sample: 0.021, 0.042, 0.022, 0.117, 0.077, 0.500, and 0.145 nM, respectively.…”