Fig. 2: Selection of nanoCLAMP binders of VHH:MTX complex. | Nature Communications

Fig. 2: Selection of nanoCLAMP binders of VHH:MTX complex.

From: Design of a methotrexate-controlled chemical dimerization system and its use in bio-electronic devices

Fig. 2

a Schematic representation of nanoCLAMP page library selection process aimed at identification of the binders selectively recognizing the VHH:MTX complex. b Inverse ELISA of recombinant purified nanoCLAMPs clones assessed for binding to VHH in the presence of indicated concentrations of MTX. Maleimide-coated plates were coated with purified nanoCLAMPs and incubated with three concentrations of biotinylated target protein and 10 μM MTX. The binding was detected using a Streptavidin–HRP-mediated colorimetric reaction. c As in (b) but without MTX. d SDS–PAGE analysis of the pulldown experiment where a mixture of biotinylated VHH domain and purified nanoCLAMP clones 1–9 were incubated with or without 10 μM of MTX and captured on the streptavidin beads prior to washing and elution with SDS–PAGE loading buffer. This experiment was performed once. Source Data are provided as a Source Data file.

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