Lu Zeng,Shanshan Song,Qiankun Zheng,Pengjie Luo,Xiaoling Wu,Hua Kuang
Shrimp tropomyosin is one of the most important causes of shellfish allergy. The objective of this study was to develop a highly sensitive and specific immunoassay for the detection of shrimp tropomyosin in food. Ten monoclonal antibodies against shrimp tropomyosin were obtained by fusion and cell screening. We found two optimum monoclonal antibodies (mAbs); mAb 2 was used as the capture antibody, and HRP-labelled mAb 3 was used as the detection antibody. Using this pair of mAbs, a sandwich enzyme-linked immunosorbent assay (ELISA) was developed with the limit of detection (LOD) of 0.45 ng/mL. Cross-reactivity with other food allergens using this method was found to be negligible. An immunochromatographic assay strip was also developed for the rapid detection of shrimp tropomyosin with the LOD of 0.5 ng/mL. The results demonstrated that our developmental methods represent a useful tool for the detection of shrimp tropomyosin in food.
Jia-Li Gu,Wen-Yan Pi,Wei-Feng Xie,Jia-Wen Gu,Ya-Jun Wang,Li Xiao,Yong-Jin Li,Jing-Fen Li
The present work is aimed to improve the antibody adsorption performance of phenyl-based mixed-mode adsorbents by adjusting the functional group of ligand. Two new mixed-mode adsorbents coupled with 4-(aminomethyl)phenol (AMP) and 4-(aminomethyl)benzonic acid (AMA) were designed based on the structure of benzylamine (BA), where a hydroxyl or carboxyl group was introduced. Bovine serum Immunoglobulin G (IgG) and bovine serum albumin (BSA) were used as the model proteins. The adsorption isotherms of IgG and BSA were investigated at different pH values ranging from 4.0–8.9 to evaluate the adsorption performance, and protein-ligand molecular interactions were explored through addition of sodium chloride and glycerol. The results showed that both of adsorbents coupled with AMP or AMA had lower saturated binding capacities ( Q m ) towards BSA and higher adsorption capacity ratio of IgG to BSA when compared to BA-based adsorbents. The contributions of electrostatic interactions and hydrophobic interactions to proteins binding were different among these phenyl-based mixed mode adsorbents. Finally, AMP-based adsorbent possessing high adsorption capacity ratio of IgG to BSA and salt tolerance was selected to separate IgG from mimetic serum with purity higher than 96 %. The results indicated adjusting the functional groups of ligand to regulate the electrostatic and hydrophobic interactions of protein-ligand was useful to improve the antibody separation performance of mixed-mode adsorbents.
危险品化学品经营许可证（带存储）