Abstract

Nanobodies® are heavy chain variable domains (VHH) derived from camelid antibodies capable of binding therapeutic targets and common protein tags with high affinity and specificity. Their desirable properties including small size, stability and solubility have been exploited in the development of novel protein affinity purification technologies. Here we describe the production of a GFP-specific affinity resin through the coupling of an anti-GFP Nanobody® to NHS-activated agarose. The single domain anti-GFP Nanobody® was expressed and purified from E. coli and tested for its ability to bind GFP and YFP, as well as any cross-reactivity towards mCherry. After conjugation the resulting resin was tested for its ability to bind GFP as well as a GFP-tagged protein target. Ultimately, it is hoped the resin can be used across automated and bespoke platforms for purification of GFP-tagged proteins to increase probability of success and decrease cycle times in the purification of challenging targets across the portfolio.