Expression of biologically active measles virus hemagglutinin glycoprotein by a recombinant baculovirus

In this study, one of the measles virus membrane proteins, named hemagglutinin (H) which has a key role in tropism, receptor binding, hemagglutinating activity and also induction of protective immunity against viral infection, was expressed by the baculovirus expression system using specific plasmid (pDONR221) to produce entry clone. Measles Virus (AIK-C strain) genome was extracted from infected Vero cells. H gene was amplified by specific primers during RT-PCR reaction and inserted into the specific plasmid (pDONR221) using BP recombination reaction. Recombinant baculovirus harboring H gene was consequently constructed by LR reaction. Insect cells (Sf9) were infected with recombinant baculovirus. In order to increase viral titer, recombinant baculoviruses were passaged four times in Sf9 cells. Synthesis of H protein was verified by SDS-PAGE, western-blot and indirect immunoflourescene using goat polyclonal antibody against Measles Virus. The results showed that H protein was partially glycosylated but it appeared to be active in hemagglutination assay. © 2008 Asian Network for Scientific Information

Microtitration of Rubella Virus in Monovalent Vaccinal Products

"nBackground: Potency test for control of rubella vaccine is a significant factor to qualify production line and vaccination pro­gram. For this reason, WHO recommends to use the microtitration method by both vaccine companies and control laborato­ries. Then the study was done to improve this test."nMethods: Three rubella virus samples, including an in-house standard, a lot of vaccine and an in- process product, were tit­tered in cell culture tubes. Then micro titration steps were tested on 96-well microplate using cocultivation of standard ru­bella vaccine dilutions and RK- 13 cell line. After 6-7 days, final reading was done and calculated the titer. Two other sam­ples were assayed with the micromethod."nResults: Titer reduction less than 0.5 log was acquired for each sample during frequent tests and between two methods."nConclusion: The procedure was profitable and accurate for potency and identity tests of rubella virus vaccine, on the basis of WHO recommendations. &nbsp