Aim: The present study aimed to investigate gingival crevicular fluid (GCF) levels of possible novel biomarkers Annexin-A1 (ANX A1), Carbonic anhydrase- 1 (CA I), and Elongation Factor-1 Gamma (EF1-Ɣ) in health along with different periodontal diseases. Material & methods: In total, 80 systemically healthy individuals were included in this study; 20 with periodontitis stage 3 grade B (P-Stage III/B), 20 with periodontitis stage 3 grade C (P-Stage III/C), 19 with gingivitis, and 21 with clinically healthy periodontium (H). Probing depth, clinical attachment level, plaque index, and papillary bleeding index were recorded. GCF ANX A1, CA I and EF1-Ɣ levels were analyzed by enzyme-linked immunosorbent assay (ELISA). Non-parametrical tests were used for statistical analysis. Results: All clinical parameters were significantly higher in periodontitis patients compared to healthy individuals (p<0.001). PStage III/B, P-Stage III/C, gingivitis and healthy groups had similar GCF ANX A1 and CA I total (p> 0.008). P-Stage III/B and P-Stage III/C had lower EF-1α total amount compared to healthy groups (p< 0.008). The gingivitis group had higher GCF EF1-Ɣ than the P-Stage III/B and healthy groups (p < 0.008). The GCF EF1-Ɣ level was a predictive protein for the diagnosis of gingivitis according to the ROC analysis (sensitivity 100%, specificity 81%, 201 cut-off value). Conclusions: The elevated GCF EF1-Ɣ levels in gingivitis imply that EF1-Ɣ may be associated with gingival inflammation. GCF EF1-Ɣ levels showed good diagnostic values, therefore may be useful to discriminate periodontitis from gingivitis and periodontal health. Further studies are needed to elucidate the role of these molecules in the pathogenesis of the periodontal disease. Keywords: gingival crevicular fluid , periodontitis, gingivitis