Single-cell cloning enables the selection of more productive Drosophila melanogaster S2 cells for recombinant protein expression.

ABSTRACT: The generation of monoclonal cell lines is an important early process development step for recombinant protein production. Although single-cell cloning is an established method in mammalian cell lines, straightforward protocols are not yet available for insect cells. We describe a new method for the generation of monoclonal insect cells without using fetal bovine serum and/or feeder cells pretreated by irradiation or exposure to mitomycin. Highly productive clones of Drosophila melanogaster S2 cells were prepared in a two-step procedure, comprising the establishment of a polyclonal population and subsequent single cell isolation by limiting dilution. Necessary growth factors were provided by co-cultivation of single transformants with untransfected feeder cells, which were later rem

SUBMITTER: Zitzmann J 

PROVIDER: S-EPMC6037645 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature