Skip to main content
. 2009 Dec 21;4(12):e8355. doi: 10.1371/journal.pone.0008355

Figure 2. Analysis of expression and binding properties of secreted zipper-tagged scFv.

Figure 2

(A) Western blot analysis of culture medium from A549 human lung carcinoma cells transduced with Ad5 vectors encoding each zipper-scFv G28-5 molecule. Lanes denote the zipper domain fused with each scFv. Culture medium was collected, Laemmli buffer added, and samples boiled prior to SDS-PAGE and western analysis with an anti-Penta-His antibody. Molecular mass markers indicate kilodaltons. (B) ELISA-based binding assays using metal affinity-purified zipper-G28-5 proteins. Each well contained 500 µg of CD40. Zipper-scFv G28-5 proteins were added to the wells at the dilutions indicated. The positive control parental G28-5 mAb was added at dilutions of 1/500, 1/2,500, 1/5,000 and 1/10,000 from a stock solution of 0.15 mg/ml.