4.2. Antibodies
celiac disease
CELIAC DISEASE
Anti-ZED1227. A mouse monoclonal antibody (clone A083, Zedira, Darmstadt, Germany) was raised against the ZED1227 analog conjugated with KLH (Keyhole Limpet Hemocyanin). Briefly, the Michael acceptor warhead was replaced by (L)-homocysteine, a non-proteinogenic thiol amino acid, thereby facilitating the nucleophilic coupling of the peptidomimetic compound to maleimide-activated KLH (Sigma, K0383, St. Louis, MO, USA). The conjugate was used to immunize mice, followed by the identification of a preferred hybridoma cell clone. The screening proved that the monoclonal IgG class antibody A083, purified by protein A chromatography, binds to ZED1227 when covalently bound to TG2 (Anti-TG2. Total transglutaminase 2 was detected with the mouse monoclonal antibody CUB7402 (Thermo Fisher, Waltham, MA, USA), which has been widely used in celiac disease immunofluorescence studies (e.g., [Anti-gliadin. Detection of gliadin peptide was done with mouse monoclonal antibody A161 (Zedira), which was raised against the synthetic deamidated gliadin-related peptide Lys57-Glu65-[α-gliadin (58-73)] (KLQPFPQPELPYPQPQ). It detects both non-deamidated (native) gliadin and deamidated alpha-gliadin 33 mer, but not gamma-gliadin 26 mer (Zedira).