원문정보
초록
영어
Baculovirus expression system is considered to have several benefits for production of recombinant protein such as high capacity, flexibility, and similar human glycosylation pattern. An accurate titering of recombinant baculovirus is required to ensure the most appropriate expression conditions in baculovirus insect cell. BaculoELISA Titer Kit (Clontech, Mountain Biew, CA, USA) is one of baculovirus titration methods. In this method, the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) type baculovirus can be measured by a standard ELISA. However, control virus of this kit applied to this method is highly expensive. In this study, expression of monoclonal antibody (mAb) CO17-1A genes by baculovirus infection was optimized at concentrations of baculovirus, passages of baculovirus and different insect cell harvesting times. Immunoblot analysis confirmed expression of mAb CO17-1A in the infected cells. mAb CO17-1A expressed significantly at the highest expression level under the conditions of 400㎕ of baculovirus passages 3(P3) in 5 ml insect cells. In addition, mAb CO17-1A expressed with a higher level when the infected cells were harvested after 72h after baculovirus infection. The results propose that baculovirus concentration, virus passage, harvesting times after virus infection conditions of can be optimized to enhance the mAb production in insect cell system without BaculoELISA assay.