Background: Thymidine kinase 1 (TK1) is a key enzyme involved in DNA synthesis. The aim is to assess the prognostic significance of serum TK1 protein concentration (STK1p) and its role in monitoring of individual customized therapy in non-small cell lung carcinoma (NSCLC) patients in routine clinical setting. Methods: A prospective study of 129 NSCLC patients was confirmed by imager, /pathology and treated by radical resection (RR) combined with individual-chemotherapy or individual-chemotherapy alone, in 2010 to 2017. The STK1p was measured using an ECL dot blot assay in sera of patients. Results: Comparisons between 2-cycle-post-individual-chemotherapy and pre-individual-chemotherapy showed that STK1p was significantly associated with treatment effect (p<0.001), and the STK1p low-group correlated significantly to the early/middle clinical stage, as well as the treatment with RR+ individual-chemotherapy (p < 0.05). A significantly poor overall survival (OS) was found in elevated-risk STK1p vs. low-risk STK1p values (p=0.016), in advantage clinical stage vs. early/middle clinical stage (p=0.004), in SCC patients compare vs. AC patients (p=0.003) and in chemotherapy alone vs. RR combined with individual-chemotherapy (p=0.001). The multivariate analysis showed that STK1p (hazard risk=2.295, p=0.010), and RR combined with individual-chemotherapy (hazard risk=3.04, p=0.0001), were independently survival factors. Conclusions: STK1p correlates significantly to survival and is an independent multivariate prognostic factor in NSCLC patients. STK1p as a low-cost assay, is a useful tool to combine with imager for a rational approach to increase the efficacy in early detection of tumor in lung cancer screening and assessment of individual adjusted therapy in NSCLC patients.