Immunochemical characterization of cc49scFv-FasL
. (A) Cytotoxicity was detected against A20 target by using culture supernatants from cc49scFv-FasLext and L6scFv-FasLext transfected BW5147 and 3T3, respectively. Culture supernatants from BW5147 and 3T3 had no detectable cytotoxicity against A20. (B and C) Membrane filtration analysis of cytotoxic materials: A 300 kDa membrane molecular filtration was used as described in Methods. Flow-through and “Retentate” of cc49scFv-FasLext (B) and L6scFv-FasLext (C) samples were readjusted to original volume and tested for cytotoxicity against A20. (D) Polyclonal rabbit anti-FasL (C-20) and rabbit anti-HA were used to probe for the presence of fusion proteins. Dilutions used were 1000 fold for anti-HA, 1000 fold for anti-C-20, and 4000 fold for horseradish peroxidase-conjugated anti-rabbit Ab.