Cell Line Generation Protocol

Unlike the short term protein expression observed using transient transfection methods generating cell lines using lentiviral vectors enables long term protein expression studies.

Cell line generation protocol.

This is usually the first step before moving to stable cell line development in order to be able to ensure that all antibody features meet the requirements in cho based productions. The protocol for generating stable cell lines requires several steps as shown below. After thawing from liquid nitrogen sw480 cells were maintained in nunclon delta t25 cell culture flasks in gibco dmem medium supplemented with 10 gibco fbs and 1 pen strep for 2 passages before seeding for spheroid generation. Turnaround time does not include waiting time for any customer supplied materials e g.

Production completion is achieved once the final cell line passes qc and is ready to be shipped out. Generate a kill curve to determine the optimal selection antibiotic concentration. For optimal results we recommend following the cell culture recommendations of the supplier. Transfect cells with desired plasmid construct s select and expand stable polyclonal colonies.

This protocol is specific for the generation of a monoclonal cell line that resistance to antibiotics g418 neomycin. Infection of human b cells with ebv in vitro results in their immortalization and the resulting cell lines are named lymphoblastoid cell lines lcls in these cells ebv establishes mainly a latent infection characterized by the expression of a limited number of viral proteins. Our xtencho tm cell line together with our in house xten protocol is your best chance at overcoming your difficult to express protein challenge. For stable cell line generation services vectorbuilder defines estimated turnaround as the time from production initiation to completion.

Cell lines or. The end result that you are looking for is a population of cells in which 100 of cells are expressing your fusion protein. This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector. Unlike the short term protein expression observed using transient transfection approaches generating cell lines using lentiviral vectors enables long term protein expression studies.

Culture conditions for generation of stable cell lines as for transient transfection experiments culture conditions passage number split rhythm etc of your selected cell type are very important for the generation of stably transfected cell lines. Epstein barr virus ebv is a lymphotropic γ herpes virus. This protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector. Identify single clones by limited dilution.