Nanobodies (the smallest antibody fragments that retain full antigen-binding capacity) are rapidly gaining traction as promising tools for pharmacologic undertakings. Conjugation of ligands with nanobodies can provide G protein-coupled receptor (GPCR) agonists with substantially greater activity and selectivity than prototype ligands from which they were derived. Biased agonism, the ability of a receptor to differentially activate downstream signaling pathways, has emerged as a powerful strategy for developing safer and more effective medications. In this work, we hypothesized that the conjugation of nanobodies to GPCR ligands could yield biased ligands. We tested this hypothesis on the neurokinin-1 receptor (NK1R), a GPCR targeted by the peptide agonists substance P (SP) and neurokinin A (NKA), which signals via multiple pathways and contributes to pain perception in humans.  We generated several NKA- and SP6-11-(a truncated version of SP) nanobody conjugates using enzymatic ligation chemistry and evaluated their signaling properties. We found that NKA conjugation to nanobodies causes prolonged cAMP production with a loss of efficacy, while Nb-SP6-11 conjugates failed to elicit cAMP. In addition, NKA and SP6-11 conjugation to nanobodies decreased β‐arrestin2 recruitment and had little impact on Gq signaling. These findings demonstrate that the conjugation of Nbs with peptide ligands for GPCRs offers a new avenue for the discovery of compounds with useful properties such as biased agonism or prolonged duration of action. Such biased ligands provide an unprecedented level of control of receptor functions, allowing for fundamental inquiries into GPCR signal transduction mechanisms. Such tools hold promise for translational efforts to develop efficacious and long-lasting therapies with reduced side effects.