Oral cancer is the leading cancer worldwide with significant morbidity and mortality rate. Use of tobacco in form of both chewing and smoking have been known as major risk factors along with alcohol and HPV infection. Despite such knowledge, there are yet no proper screening strategy employed for screening of oral cancer at early stage. In this study, we employed TMT-based comparison of proteins from serum samples from healthy volunteers and OSCC patients with chewing and smoking habits. The data has been acquired in triplicate where we have used multiple fractionation strategies such as bRPLC and SCX-based fractionation. The candidate proteins identified at discovery phase were further validated using ELISA.