Fig. S1.

Selection and purification of human anti-NCL scFvs by phage display. (A) Binding of selected phage clones or soluble scFvs to NCL was assessed by ELISA using NCL-coated plates incubated with the indicated clones. Clone 4LB5 used for further experiments is indicated with an asterisk (*). The assay was performed three times in triplicate using different preparation of phages and scFvs, and mean ± SD is reported. (B) Selected clone named 4LB5 was subcloned in pET22b E. coli expression vector and transformed in BL21-DE3 bacterial cells. (C) IPTG-induced scFv was extracted from inclusion bodies, then refolded using urea gradient, and finally purified using Ni-NTA columns. FT, flow through; M, molecular standard; 1–6, Different elutions of the scFV.