Purpose <p>Pancreatic ductal adenocarcinoma (PDAC) is the most prevalent form of pancreatic cancer, with high malignancy and poor prognosis. The cellular mesenchymal-epithelial transition factor (c-Met) is overexpressed in 84% of PDAC and plays a critical role in tumor progression, which is closely associated with poor patient outcomes. In this study, a new ⁶⁸Ga-labeled nanobody ([⁶⁸Ga]Ga-NOTA-PFCM01) was developed for the visualization of c-Met in PDAC models.</p> Methods <p>The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) data were utilized to assess <i>MET</i> expression and overall survival in patients with different cancers. By immunizing an alpaca with recombinant human c-Met, three clones of nanobodies were screened, and the binding affinity was tested by bio-layer interferometry (BLI). The binding epitope of the nanobodies and c-Met was predicted by AlphaFold3. c-Met expression in human PDAC cell lines was evaluated using western blot, flow cytometry, and confocal microscopy. NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) chelator was used to label the nanobodies with ⁶⁸Ga. PET imaging, semiquantitative uptake, and biodistribution research were carried out in tumor xenografts. Histological staining was performed on tumor tissues to characterize the expression of c-Met. In addition, PET imaging in non-human primates was performed to assess pharmacokinetics and biodistribution of the nanobody.</p> Results <p>Based on TCGA and GTEx data, <i>MET</i> expression in pancreatic adenocarcinoma (PAAD) is significantly higher than that in normal tissues (<i>P</i> &lt; 0.001). Patients with high <i>MET</i> expression have lower overall survival rates than those with low <i>MET</i> expression. c-Met expression was the highest in BXPC-3 cells but the lowest in MIA PaCa-2 cells, which were set as the positive and negative models respectively. PFCM01 was screened and selected with an excellent binding property with the <i>K</i>_<i>D</i> value of 0.16 nM. The radiochemical purity (RCP) of [⁶⁸Ga]Ga-NOTA-PFCM01 was high, with good in vivo and in vitro stability. Semiquantitative analysis of small animal positron emission tomography/computed tomography (PET/CT) imaging demonstrated significantly higher tumor uptake in BXPC-3 tumors at 2&#xa0;h post-injection (2.31 ± 0.02%ID/g) than control groups. Ex vivo biodistribution also showed a high uptake of BXPC-3 tumors (1.90 ± 0.59%ID/g) than others, which was further verified the PET imaging results. Histological staining showed high expression of c-Met in BXPC-3 but low in MIA PaCa-2 tumor tissues. In healthy cynomolgus monkeys, PET imaging revealed rapid renal excretion of [⁶⁸Ga]Ga-NOTA-PFCM01. No significant radioactive uptake was observed in the liver, stomach, intestines, pancreas, or muscles, indicating its favorable pharmacokinetics and translational potential.</p> Conclusions <p>[⁶⁸Ga]Ga-NOTA-PFCM01 showed a specific and high tumor uptake in c-Met-positive PDAC models, providing a noninvasive method to assess c-Met expression. Besides, it demonstrated good pharmacokinetic characteristics in non-human primates. Therefore, [⁶⁸Ga]Ga-NOTA-PFCM01 provided the potential role as a prospective PET imaging agent for future clinical applications.</p>