Analysis of scFv clones by agarose gel electrophoresis (1%) and purification of scFvs. A. Agarose gel analysis of colony PCR of 10 randomly selected clones from library before panning. B. Agarose gel analysis of SfiI and NotI digested plasmid of 10 randomly selected clones from library before panning. Lane M, 100 bp marker; Lane M1 1kb ladder; Lane 1–10 are plasmid DNA from ten different clones. C. SDS- PAGE (12 %) of purified scFv Lane M, protein marker; Lane E1 to E5, different eluted fractions of purified scFv. Arrow indicates the 32kDa band of scFv in E1 to E5 lanes. D. Western blot analysis of purified scFv, pCANTAB-5E vector periplasmic extract was used as a negative control. Lane M, prestained molecular weight marker.
Kumar et al. BMC Biotechnology 2012 12:87 doi:10.1186/1472-6750-12-87