Dictyophora rubrovalvata is a valuable fungus homologous to food and medicine,and its polysaccharide have been gaining increasing attention because of its plentiful activity.However,the structure and activity of its h...Dictyophora rubrovalvata is a valuable fungus homologous to food and medicine,and its polysaccharide have been gaining increasing attention because of its plentiful activity.However,the structure and activity of its homogeneous polysaccharide have not been studied enough.In this study,two polysaccharides DRP-I and DRP-II were purified from D.rubrovalvata.Their structures were characterized by chemical composition,monosaccharide composition analysis,methylation analysis and nuclear magnetic resonance spectroscopy.The results showed that DRP-I and DRP-II were neutral heteropolysaccharides with molecular weights of 5.79×103 and 1.25×104 Da,respectively,which were composed of mannose,galactose,glucose,xylose and fucose.The main chains were→6)-α-D-Galp-(1→6)-α-D-Galp-(2,1→6)-α-D-Manp-(2,1→6)-α-D-Galp-(1,and branch chains wereβ-D-Xylp-(1→3)-α-L-Fucp-(1→4)-α-D-Manp-(1→andα-D-Galp-(1→3)-α-D-Galp-(1→.The in vitro immunoactivity assays on dendritic cells showed that DRP-I and DRP-II could up-regulate the expression of IL-10 and IL-6 and inhibit the expression of TNF-αin a concentration-dependent manner.This research indicated that DRP-I and DRP-II possessed immunoactivity by balancing the excessive inflammation,and molecular weight is an important factor affecting immunoactivity.展开更多
To purify Methamidophos (Met) monoclonal antibodies with two methods and compare immune activity of purified antibodies. Method Caprylic acid ammonium sulphate precipition (CAASP) method and Sepharose protein-A (S...To purify Methamidophos (Met) monoclonal antibodies with two methods and compare immune activity of purified antibodies. Method Caprylic acid ammonium sulphate precipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method were used to purify Met monoclonal antibodies, UV spectrum scanning was used to determine protein content and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linked immunosorbent assay (ELISA) was used to determine immune activity of purified antibodies. Results Antibody protein content and recovery rate with CAASP method were 7.62 mg/mL and 8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mL and 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASP method. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was 181.26 靏/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01 靏/mL and 1.03 靏/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82 靏/mL, and the linear working range and LOD were 10.91-11412.29 靏/mL and 3.42 靏/mL, respectively. Conclusion Antibodies purified by SPA method are better than those by CAASP method, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelled testing paper for analyzing Met residue in vegetable and drink water.展开更多
基金Key Project of Yunnan Provincial Nature Foundation(202401AS070075)。
文摘Dictyophora rubrovalvata is a valuable fungus homologous to food and medicine,and its polysaccharide have been gaining increasing attention because of its plentiful activity.However,the structure and activity of its homogeneous polysaccharide have not been studied enough.In this study,two polysaccharides DRP-I and DRP-II were purified from D.rubrovalvata.Their structures were characterized by chemical composition,monosaccharide composition analysis,methylation analysis and nuclear magnetic resonance spectroscopy.The results showed that DRP-I and DRP-II were neutral heteropolysaccharides with molecular weights of 5.79×103 and 1.25×104 Da,respectively,which were composed of mannose,galactose,glucose,xylose and fucose.The main chains were→6)-α-D-Galp-(1→6)-α-D-Galp-(2,1→6)-α-D-Manp-(2,1→6)-α-D-Galp-(1,and branch chains wereβ-D-Xylp-(1→3)-α-L-Fucp-(1→4)-α-D-Manp-(1→andα-D-Galp-(1→3)-α-D-Galp-(1→.The in vitro immunoactivity assays on dendritic cells showed that DRP-I and DRP-II could up-regulate the expression of IL-10 and IL-6 and inhibit the expression of TNF-αin a concentration-dependent manner.This research indicated that DRP-I and DRP-II possessed immunoactivity by balancing the excessive inflammation,and molecular weight is an important factor affecting immunoactivity.
基金National Natural Science Foundation of China (39970519) Guangxi Basic Studying Fund (0236053) +1 种基金 Guangdong Province Natural Science Foundation (990683) Core Teacher Project of Ministry of Education and Chinese Postdoctoral Fu
文摘To purify Methamidophos (Met) monoclonal antibodies with two methods and compare immune activity of purified antibodies. Method Caprylic acid ammonium sulphate precipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method were used to purify Met monoclonal antibodies, UV spectrum scanning was used to determine protein content and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linked immunosorbent assay (ELISA) was used to determine immune activity of purified antibodies. Results Antibody protein content and recovery rate with CAASP method were 7.62 mg/mL and 8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mL and 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASP method. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was 181.26 靏/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01 靏/mL and 1.03 靏/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82 靏/mL, and the linear working range and LOD were 10.91-11412.29 靏/mL and 3.42 靏/mL, respectively. Conclusion Antibodies purified by SPA method are better than those by CAASP method, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelled testing paper for analyzing Met residue in vegetable and drink water.
