Gluten tespitindeki ilerlemeler: Antikor gelişimi ve yanal akışlı immünodeney için altın ​​nanopartikül konjugasyonu

Objective: Gluten-related disorders, like celiac disease (CD), affect 5% of the global population, with CD impacting 1.4% of individuals. Consuming gluten triggers inflammation in the intestines for those with CD. Following a gluten-free (GF) diet is crucial despite challenges like cross-contamination. Immunoassays like enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay (LFIA) are preferred for detecting gluten due to their simplicity and cost-effectiveness. LFIA kits offer a portable option for analyzing gluten in food. The demand for reliable gluten detection methods highlights the need for accurate solutions for both food manufacturers and individuals with gluten-related disorders. Our research focuses on developing specific techniques using antibodies targeting gliadin, one of the main parts of gluten, to confirm the absence of gluten. Method: The polyclonal antibodies obtained from rabbits were purified and tested for reactivity to gliadin using ELISA and Western blot techniques. They were then conjugated with gold nanoparticles (AuNPs) to develop an LFIA, with optimal conditions determined by pH and antibody concentration. The efficacy of the LFIA in detecting wheat gliadin, including the limit of detection, was investigated. Gliadin was extracted and analyzed from various food samples to establish a reliable method for detecting it in different products. Results: Antibodies from three rabbits (R1, R2, and R3) were sensitive to gliadin in an ELISA. Western blot analysis confirmed common gliadin-specific epitopes in these antibodies. They did not react to GF food samples. The LFIA effectively detected wheat gliadin using a selected membrane and fibers, with a detection limit of 50 µg/mL. The LFIA consistently produced reliable results when testing samples from different food sources, making it a dependable tool for gliadin detection in various food samples. Conclusion: Our study presents a technique for identifying gluten, including the development of antibodies, improving AuNP attachment, and establishing a reliable LFIA protocol. This methodology has the potential to ensure food safety and facilitate gluten analysis. Keywords: Antibody development, Antibody conjugation, Lateral flow immunoassay