Recombinant antibodies, scFv-Fc and scFv, derived from anti-IGF-IR monoclonal antibody 1H7, were previously expressed in mammalian NSO and E.coli cells, respectively [Li et al. Cancer Immunol. Immunother. 49, 243, 2000; Kusada et al. J. Biochem. 143, 9, 2008]. Expression of 1H7 scFv proteins in E. coli, however,did not yield significant quantities. To achieve better yields, we used a Drosophila S2 cell expression system to express 1H7 scFv. The 1H7 scFv gene cloned into a pMT/BiP/V5-His expression vector was co-transfected with a pCoHygro selection vector into S2 cells. After 4-weeks culture in the presence of 0.3 mg/ml hygromycin B, stably-transfected cells were treated with 0.5 mM CuSO4 to induce scFv protein expression. From culture supernatants, 1H7 scFv proteins were partially purified by Ni2+-Sepharose chromatography. IGF-IR binding activity measured using SPR and ELISA revealed production of active 1H7 scFv proteins. Alternative protocols are being tested to improve purification purity and yield.

Purification of anti-insulin-like growth factor-I receptor (IGF-IR) single chain antibody (scFv) expressed in <i>Drosophila</i> S2 cells

244th National Meeting (2012)

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Purification of anti-insulin-like growth factor-I receptor (IGF-IR) single chain antibody (scFv) expressed in Drosophila S2 cells