Isolation of NCAM1-binding scFvs by phage display.

<p>(A) Titres of input and output phage populations throughout library panning. Pfu = phage-forming units. (B) ELISA analysis of NCAM1-Ig1 binding activity of eluted polyclonal phage-scFv populations. P0: Unpanned library. P1-P4: Polyclonal phage preparations eluted after rounds 1 to 4 of library panning against immobilised NCAM1-Ig1. NCAM1-Ig1 was coated in ELISA wells at 100 µg/ml. Bars represent the mean of three replicate wells and error bars indicate the standard deviation of the mean. (C) Competitive ELISA analysis of NCAM1-Ig1 binding of B5 scFv. Binding of the purified scFv to immobilised NCAM1-Ig1 (100 µg/ml) was out-competed by pre-incubating the scFv with soluble NCAM1-Ig1 domain prior to ELISA analysis. SMP: skimmed milk powder control. Bars represent the mean of three replicate wells and error bars indicate the standard deviation of the mean.