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. 2004 Jun;14(6):1119–1129. doi: 10.1101/gr.1821804

Figure 5.

Figure 5

(A) Soluble scFv Western. (Lanes 1,2) D1.3 scFv expressed from a control vector that produces scFv with attached SV5 and 6x His tags at the carboxyl terminus; (lanes 3,4) D1.3 scFv expressed from the pD1.3 destination vector that produces the D1.3 scFv with FLAG and translated Lox 2372 at the amino terminus and Lox WT, SV5, and 6x His tags at the carboxyl terminus. A total of 10 μL of supernatant was used in lanes 1 and 3, and 2 μL in lanes 2 and 4 (see Methods). Arrows denote correctly sized scFv visualized with anti-SV5 and goat anti-mouse-AP conjugate antibodies. (B) scFv ELISA was performed with pDes 3A and F4 against SNV-N, E-coil GFP, and K-coil GFP coated on a 96-well plate. Antigen reactive scFv were detected by anti-SV5 tag antibody followed by anti-mouse antibody conjugated to horse radish peroxidase. (C) scFv ELISA was performed with pDes 3A E-coil scFv incubated with K-coil alkaline phosphatase (K-AP) against SNV-N and lysozyme. Control reactions measured K-coil reactivity toward SNV-N, lysozyme, E-coil GFP, and pDes 3A-E scFv.