Production of anti-dengue NS1 monoclonal antibodies by DNA immunization

Monoclonal antibodies against dengue NS1 protein were generated following immunization of mice with plasmid DNA encoding the transmembrane form of NS1 from dengue serotype 2 virus. A mammalian expression vector, pDisplay, was engineered to direct cell surface expression of dengue NS1 and tested for transient expression in COS cells. Two mice were immunized intramuscularly with six doses of 100 microg of plasmid at 2-week intervals; one mouse received a booster of live virus prior to the last plasmid injection. Both mice showed antibody responses against dengue antigens in dot enzyme immunoassay. Following fusion, hybridomas were screened with dot enzyme immunoassay against all four dengue serotypes. Specificity to the NS1 protein was confirmed by western blot analysis. Among five anti-dengue NS1 monoclonal antibodies generated, two clones were serotype 2 specific, two clones reacted with all four serotypes and the last also reacted with Japanese encephalitis virus. Reactivity against native or denatured forms of NS1 revealed three clones with reactivity to linear epitopes and two clones recognizing conformational epitopes. Such diverse specificity of anti-dengue NS1 monoclonal antibodies indicates that DNA immunization, especially with the combination of virus boosting, is an efficient way of producing monoclonal antibodies against viral protein. This has opened up a possibility of producing monoclonal antibodies to rare viral proteins that are difficult to isolate or purify.     

Preparation of activated carbon from Dipterocarpus alatus fruit and its application for methylene blue adsorption

Activated carbons were prepared from three parts of Dipterocarpus alatus fruit (wing, endocarp and pericarp), an abundant and renewable waste in Southeast Asia, by chemical activation using ZnCl₂, FeCl₃, H₃PO₄ and KOH and physical activation using CO₂ and steam. This study indicated that activated carbon prepared from Dipterocarpus alatus fruit could be employed as a promising adsorbent for the removal of methylene blue from aqueous solution. ZnCl₂ activation led to an activated carbon with a surface area of 843 m² g⁻¹ and was able to remove methylene blue from aqueous solution. Adsorption studies were performed and analysed using Langmuir and Freundlich isotherm equations. Adsorption data demonstrated an excellent fit with the Langmuir isotherm model, with the maximum adsorption capacity of 269.3 mg g⁻¹ at equilibrium. Pseudo-first order and pseudo-second order kinetic models were used in this study to describe the adsorption mechanism. The results show that methylene blue adsorption is pseudo-second order, indicating that liquid film diffusion, intra-particle diffusion and surface adsorption coexisted during methylene blue adsorption on the activated carbon. The activated carbon prepared from Dipterocarpus alatus fruit is a low cost and effective adsorbent with a fast rate for the removal of methylene blue from aqueous solutions when compared with a number of activated carbons studied in the literature.

Activated carbons were prepared from Dipterocarpus alatus fruit by chemical and physical activation and used for the removal of methylene blue from aqueous solution.