Production of calmodulin-tagged proteins in Drosophila Schneider S2 cells: a novel system for antigen production and phage antibody isolation.

We report the development of an expression system for the production of soluble, calmodulin (CaM)-tagged proteins in Drosophila Schneider S2 cells and the subsequent use of these proteins for the selection of phage displayed antibodies. The CaM-tag permitted the purification of recombinant protein to >90% purity in a single step at yields of >20 mg/l. Using platelet glycoprotein VI (GP6) as a model, we demonstrated that the recombinant CaM-tagged protein was post-translationally N-glycosylated and had identical ligand specificity to native protein. A novel selection strategy, exploiting the CaM tag, was then used to isolate four single chain Fv fragments (scFvs) specific for GP6 from a non-immune phage display library. In contrast to other selection methods, which can result in antibodies that do not recognise native protein, all of the scFvs we selected bound cell surface expressed GP6. In conclusion, the production of CaM-tagged proteins in Drosophila Schneider S2 cells and the selection strategy reported here offer advantages over previously published methods, including simple culture conditions, rapid protein purification, specific elution of phage antibodies and preferential selection of phage antibodies that recognise native, cell surface expressed protein

First fossil finds of some Australian Bryozoa (Cheilostomata)

The abundant fossil record of well-preserved Bryozoa in samples from the Tertiary of Victoria and South Australia includes some \u27first fossil finds\u27 which are recorded here. Several are of species known from the Recent of the Australian or Indo-West-Pacific regions, but some represent genera with a much wider temporal and geographical range. Of the 11 species illustrated, six are known, or may be inferred, to have inhabited \u27sand fauna\u27 environments. Specimens of one species are complete enough to allow its formal description as Chlidoniopsis inopina sp. nov. <br /