INTRODUCTION: Parkinson's disease (PD) is a progressive neurodegenerative disease, and biomarkers are needed to enhance earlier detection and monitoring. Alpha-synuclein, phosphorylated at serine 129 (pS129-α-syn), is the predominant form of α-syn found in Lewy bodies implicating an involvement in disease pathology. This review aims to systematically evaluate the evidence for pS129-α-syn detection in human biofluid samples of PD utilizing ELISA-based protein detection methods. METHODS: A systematic review was conducted following the Preferred Reported Items for Systematic Review and Meta-Analyses (PRISMA) guidelines. Electronic searches were performed in PubMed, Web of Science, and Cochrane databases from inception to November 7th, 2024, to identify studies comparing pS129-α-syn in biofluids of PD patients with controls or related neurodegenerative disease. Risk of bias was assessed for each study. RESULTS: Twenty-three publications met the inclusion criteria, with pS129-α-syn detected in cerebrospinal fluid, plasma, red blood cells, serum, and saliva exosomes. Overall, pS129-α-syn levels were elevated in patients with PD compared to controls, and in some studies, correlated with disease severity. There was no consistent pattern when comparing PD patients to those with related neurodegenerative diseases. Significant variability in pS129-α-syn levels and considerable overlap between groups may limit the utility as a biomarker. CONCLUSION: While pS129-α-syn for PD shows some promise as a diagnostic marker for PD, its differential diagnostic utility remains limited. Further research involving larger cohorts is required. The greatest potential for pS129-α-syn may be as part of a panel with other PD-specific markers, to enhance diagnostic accuracy and prognostic value.