Research on the Mechanism of Action of a Citrinin and Anti-Citrinin Antibody Based on Mimotope X27

Abstract

Immunoassays are developed based on antigen–antibody interactions. A mimotope is an effective recognition receptor used to study the mechanism of action of antigens and antibodies, and is used for improving the sensitivity of the antibody. In this study, we built a 3D structure of the citrinin (CIT) mimotope X27 and anti-CIT single-chain antibody fragment (ScFv) through a “homologous modeling” strategy. Then, CIT and X27 were respectively docked to anti-CIT ScFv by using the “molecular docking” program. Finally, T28, F29, N30, R31, and Y32 were confirmed as the key binding sites in X27. Furthermore, the result of the phage-ELISA showed that the mutational phage lost the binding activity to the anti-CIT ScFv when the five amino acids were mutated to “alanine”, thereby proving the correctness of the molecular docking model. Lastly, a site-directed saturation strategy was adopted for the sites (T28, F29, N30, R31, and Y32). Eighteen different amino acids were introduced to each site on average. The activities of all mutants were identified by indirect competitive ELISA. The sensitivities of mutants T28F, T28I, F29I, F29V, N30T, and N30V were 1.83-, 1.37-, 1.70-, 2.96-, 1.31-, and 2.01-fold higher than that of the wild-type, respectively. In conclusion, the binding model between the CIT and antibody was elaborated for the first time based on the mimotope method, thereby presenting another strategy for improving the sensitivity of citrinin detection in immunoassays.

Classification of drugs based on mechanism of action using machine learning techniques

“…Aspirin-like medications hindered the generation of physiologically significant Prostaglandins by blocking this step in the Prostaglandin production process. This gave a unified explanation for the aspirin-like medicines' therapeutic activity and its common adverse effects Research on the Mechanism of Action of a Citrinin and Anti-Citrinin Antibody Based on Mimotope X27 [8] A mimotope is a powerful recognition receptor that can be utilised to investigate antigen and antibody mechanisms of action. The mimotope method was used to develop a binding model between citrinin and antibody.…”