Developing radiopharmaceuticals by incorporating biomolecules is a powerful tool for the targeted delivery of radioisotopes. The positron-emitting radionuclide copper-64 can be complexed with the NOTA chelator to generate tracers for positron emission tomography (PET) imaging and disease diagnosis. Fibroblast activation protein (FAP) is overexpressed on cancer-associated fibroblasts, emerging as a target for PET imaging and radiopharmaceutical therapy. Nanobodies, single-domain antibodies derived from camelids, possess high affinity, small size, and excellent tissue penetration, making them ideal targeting biomolecules. Here, we use sortase-mediated enzymatic conjugation in combination with click chemistry. These techniques enable site-specific attachment of a chelator and controlled incorporation of a PEG pharmacokinetic modulator to a FAP-targeting nanobody. Following this, the tracer can be radiolabelled with copper-64 for evaluation in a FAP tumour mouse model. This strategy presents potential to generate FAP-targeted PET imaging agents.